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via Duane Jensen at https://www.youtube.com/watch?v=tJxFzACiISM&lc=UgyDGDDU6cEskMvsPmh4AaABAg.9Vb0zueAXOFAEfXxtmCuPo

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Reply to the reviewers

Response to the points raised by the reviewers.

Once again, we would like to thank the reviewers for their comments. We have systematically addressed their concerns, as detailed below.

Reviewer #1

Evidence, reproducibility and clarity

This study demonstrates that BICD2, previously known as an adaptor protein for dynein, is involved in regulating centriole engagement during mitosis. First, using different antibodies, it was shown that BICD2 localizes near the mother centriole, as observed by super-resolution microscopy. During G1 and S phases, BICD2 localizes slightly outside the Cep152 ring, while in G2 to mitosis, it localizes near the cartwheel component SAS-6. Moreover, analysis of deletion mutants revealed that BICD2 localizes to the centrosome in a CC domain-dependent manner at the C-terminal end. The localization pattern resembling a ring in the cytoplasm was also observed through the CC3 domain. Next, BICD2 knockout (KO) cells were generated to investigate centriole dynamics. In BICD2 KO cells, the distance between the mother and daughter centrioles was observed to increase from G2 to mitosis compared to controls. Along with this, early centriole disengagement and centriole amplification phenotypes were observed. The increased distance phenotype between centrioles was rescued in BICD2 wild-type (WT) and mutant forms lacking the CC1 domain at the N-terminus, suggesting that this function of BICD2 is independent of dynein. Additionally, BICD2 mutants mimicking phosphorylation at the C-terminus showed reduced centrosome localization and were unable to rescue the phenotypes seen in BICD2 KO cells.

While the study clearly demonstrates BICD2's contribution to centriole engagement, the underlying mechanisms of how BICD2 is involved in centrosome localization and centriole engagement remain unclear. As it is anticipated that the function of BICD2 is independent of dynein, further exploration of this unknown mechanism would enhance the value of the paper. Below are the concerns that should be addressed, including new experiments.

Main Points:

1. __ Fig. 1-3: Regarding the localization of BICD2 to centrioles, during the G1-S phase, its localization appears to overlap with PCM. Experimental investigation should be performed to examine whether BICD2's centrosomal localization is influenced by knockdown of PCM components like PCNT, Cep192, or Cep152.__ We now show that BICD2 localization does not depend on pericentrin (Supplementary Figure S4B). We also show that the two proteins do not colocalize (Supplementary Figure S4A and S4D) and are functionally independent (Figure 5).

We also show that BICD2 localization does depend on the torus protein CEP152 (Figure 8B). Importantly, our data indicate that BICD2 interacts with the N-terminal region of CEP152, suggesting that this interaction places BICD2 at the outer region of the torus (Figures 8C and 8D). We propose that this provides a mechanistic basis for BICD2 function in maintaining mother-daughter centriole engagement.

__ Fig. 7: The experiments using BICD2 mutants suggest that the function of BICD2 here is independent of dynein. To further investigate whether BICD2's role in centriole engagement is independent of dynein, experiments should be conducted to examine the effect of dynein knockdown on BICD2 localization to the centrosome and centriole engagement.__

Using Dynapyrazole-A, a fast-acting and potent dynein inhibitor, we demonstrate that acute inhibition of dynein motor activity affects neither the centrosomal localization of BICD2 during G2 and M phases (Supplementary Figure S3A) nor centriole engagement (Supplementary Figure S3B). Together with experiments using BICD2 mutants deficient in dynein interaction (Figure 7B), these data compellingly demonstrate that the recruitment and function of BICD2 at the centriole is dynein-independent.

__ Fig. 4: The CC4 domain at the C-terminus of BICD2 is important for its centrosomal localization, but identifying the binder/recruiter responsible for BICD2's centrosome localization would be desirable.__

We thank the reviewer for prompting us to investigate this further. We are delighted that we now identify the torus protein CEP152 as the BICD2 binder/recruiter at the centriole (Figure 8). As we discuss in the manuscript, our observation that the outward-facing N-terminus of CEP152 interacts with the C-terminal region of BICD2 provides a mechanistic basis for understanding BICD2's role in maintaining mother–daughter centriole engagement.

__ Fig. 7: Rescue experiments using BICD2 mutants suggest that BICD2's functional domains are critical. Further experiments by creating mutants missing parts of CC2 or CC3 could identify functionally important domains of BICD2 by observing any loss-of-function phenotypes at the centrosome.__

We fully appreciate the reviewer’s suggestion to examine the roles of the CC2 and CC3 regions. We believe that these domains, and particularly the unstructured loop within CC3, are important for both BICD2 localization, function and regulation at the centrosome. However, given that our current data already establish a clear mechanism for BICD2 centriolar recruitment via CEP152 and the CC4 region, we feel these additional structural studies fall outside the core scope of the present manuscript. We hope the reviewer agrees that the current evidence provides a robust foundation for our conclusions, and we look forward to addressing the roles of CC2 and CC3 in a dedicated future study.

__ Fig. 6: Regarding the BICD2 KO cell phenotype, is there experimental evidence showing an increase in centriole number during mitosis? For instance, while no abnormality in centriole number may occur during G2, a trend of increase in mitosis should be experimentally demonstrated. Also, how should the slight differences in phenotypes between Ndelta4 and Ndelta5 BICD2 KO cells be interpreted?__

We thank the reviewer for highlighting this point, but we would like to clarify that we do indeed observe a significant increase in centriole number during both mitosis and G2 phase across multiple cell lines in our BICD2 KO models and RNAi experiments (Figure 4E, RPE-1 KO cells, and 4G, U2OS cells, RNAi) and G2 (Figure 5C, both RPE-1 and U2OS, RNAi). As the main text was not explicit enough on this point, we have revised the manuscript to describe these observations more clearly.

Regarding the phenotypic differences between BICD2 KO lines, we assign them to the clone-to-clone functional heterogeneity often seen in CRISPR/Cas9-generated cell lines. Importantly both clones show a consistent, statistically significant phenotype (e.g., impaired engagement and increased centriole numbers) compared to wild-type controls, confirming that the overall defect is robust and specific to BICD2 loss. We have added a clarifying note on this in the revised manuscript: “Figure 4F; we assign the differences between BICD2-/- cell lines to standard clone-to-clone phenotypic heterogeneity often seen in CRISPR/Cas9-generated cell lines.”

__ Fig. 8: Regarding the phosphorylation of BICD2 at the C-terminus: The phenotypes of mutants where these two phosphorylation sites are changed to alanine should be experimentally observed. It is expected that the removal of BICD2 from the centrosome during mitosis could be rescued. Additionally, the effect of PLK1 or CDK1 inhibitors on the removal of BICD2 from the centrosome should be investigated.__

We agree with the reviewer that phosphonull mutants should be added to these experiments. As mentioned above we have decided to remove the preliminary data regarding BICD2 phosphorylation from the manuscript data to present a more comprehensive, dedicated study on BICD2 phosphorylation in the near future. In fact, we have already performed the suggested experiments, including the phosphonull mutants and kinase inhibitor treatments, and would be glad to share these additional results if the reviewers would find them helpful. Interestingly, our experiments show that BICD2 centrosomal amounts are not affected by PLK1 inhibition (using BI 2536); CDK1 inhibition (RO-3306), although not significatively changing the amount of BICD2 at the centrosome, slightly diminishes it. We currently favor a model in which BICD2 is predominantly regulated by CDK1, and we are actively defining the precise molecular mechanism governing this regulation.

Minor Points:

__ Fig. 1-3: During G1 and S phases, BICD2 localizes near the mother centriole, and from G2 onward, it colocalizes with SAS-6. How can this be explained?__

We currently do not have a clear explanation for this transition, as our focus has been in understanding BICD2 recruitment to the centriole (and its role in centriole engagement). We view this as a very interesting question that could be studied together with BICD2 regulation through phosphorylation. Our current hypothesis is that most of BICD2 is removed through phosphorylation in late G2 and M, with a pool remaining at the mother-daughter interface, possibly protected by a yet to be understood mechanism. We have added a sentence in the discussion addressing this (“ A pool of protein could be protected and correspond to the observed remnant of BICD2 at the mother-daughter interface.“). This last pool, as we discuss in the manuscript, could be further phosphorylated at the M/G1 transition or cleaved by separase (although this last point is of course highly speculative).

__ Fig. 4: The GFP-BICD2 488-820 fragment forms cytoplasmic rings, which is interesting. This domain contains the CC4 domain, so it can localize near the centriole, but why does it not form a perfect ring there? Also, which other centriole/centrosome markers were used for colocalization studies? Does knockdown of PCM1 affect BICD2's centrosomal localization?__

We show in Figures 6G and 6H that BICD2 488-820 can form a ring around the centriole. Indeed this polypeptide contains the CC4 region, which our results indicate it will guide it to the centriole (through an interaction with CEP152). Once the available CEP152 is occupied with BICD2 we assume that BICD2 488-820 forms oligomers that assemble ring-like structures outside the centriole.

Other centrosomal markers used are SAS-6.

We now show that PCM1 knockdown does not affect BICD2's centrosomal localization (Supplementary Figure S4C). Although our results indicate that partial forms of BICD2 such as BICD2 488-820 can colocalize with PCM-1 (Figure 6D), full length endogenous BICD2 (or GST-BICD2) does not seem to colocalize with this protein and thus the centriole satellites (Supplementary Figure S4A). We note this discrepancy in the text: “The presence of BICD2 at the centriolar satellites has been suggested previously (Quarantotti et al, 2019); we ignore the reason why in the conditions used in this study only C-terminal fragments of BICD2 but not the full-length protein”. The relationship between satellites and BICD2 grants further studies. Our data suggests that centriolar localization of the protein may be regulated, possibly by its intramolecular structure, and that regulated binding of BICD2 to a yet to be identified partner may recruit the protein to satellites either for its transport to the centrosome or in order to perform a specific function at the satellites. We have added a sentence to the text to note this: “This suggests that BICD2 satellite localization is regulated (possibly via intramolecular autoinhibition) to mediate BICD2 transport or a distinct satellite-specific function of this protein.”.

__ Fig. 4A: What are the aggregates observed in the cytoplasm under the GFP-BICD2 + ice condition? Also, does the 1-575 mutant fail to localize to the centrosome upon ice treatment?__

We currently do not know the nature of the GFP-BICD2 full length aggregates observed upon microtubule depolymerization. We also observe GFP-BICD2 aggregates in cells that express high amounts of the polypeptide, leading us to hypothesize that it may be insoluble and the disappearance of microtubules may liberate it from motor complexes resulting in its aggregation -although of course more work would be needed to clarify this.

We now show new data (Supplementary Figure S5), showing that the localization of not only GFP-BICD2 1-575 but also the C-terminal fragments 272-820 and 488-820 are not significantly affected by cold-induced microtubule depolymerization. These last results strongly suggest that BICD2 localization at the centriole is microtubule independent and are compatible with our data showing that BICD2 can directly interact with the centriolar protein CEP152.

__ Can similar phenotypes be observed in other cell types when BICD2 is knocked down? This should be experimentally validated.__

Our current manuscript now shows that similar phenotypes regarding centriole separation and amplification are observed upon BICD2 depletion in RPE-1 cells (non-transformed, p53-wildtype) and U2OS cells (transformed). These are shown in Figure 4 (RPE-1 knockout, U2OS RNAi knockdown) and Figure 5 (RPE-1 and U2OS RNAi knockdown).

__ Are there previous studies suggesting that this function of BICD2 is evolutionarily conserved? This should be addressed.__

To our knowledge there are no previous studies describing BICD2 function at the centrosome, excepting the recent article by Kuang et al., (Kuang W et al. 2025. BICD2 promotes ciliogenesis by facilitating CP110 removal from the mother centriole. EMBO reports 26:5567–5588. DOI: https://doi.org/10.1038/s44319-025-00597-0), that describes a role for BICD2 during ciliogenesis in non-cycling cells. As we mention in our discussion this new role may be related to the distal pool of protein that we observe using ExM, and we don’t think is related to the function of the proximal pool of BICD2 at the torus in cycling cells that we describe in our manuscript.

Regarding functional conservation, BICD2 orthologs are widely distributed across metazoans (as reflected in OrthoDB, which lists ~5,000 ortholog genes across ~2,500 species). They share a remarkably conserved C-terminal domain that acts as a docking interface mediating subcellular targeting independently of dynein motor activity (i.e. through binding to Rab6, RanBP2 and, as shown here, CEP152). Cross-species analyses show that this C-terminal domain is preserved in most eukaryotic orthologs, including Drosophila melanogaster BICD (UniProt P16568) and Caenorhabditis elegans BICD-1 (UniProt V6CJ04). Interestingly, several predicted orthologous sequences in public databases retain high C-terminal similarity while completely lacking the N-terminal regions containing the CC1 box motif (residues 29–57 in human BICD2) required for dynein interaction (e.g., predicted isoforms in mouse or camels). Thus, dynein-independent scaffolding functions may represent an ancient, foundational role of the BICD protein family, or alternatively (and perhaps most probably, given that basal metazoans like sponges or Cnidaria do show a conserved N-terminus), these truncated forms may have evolved to fulfill distinct cellular roles operating independently of motor-adaptor activity. We have added a passage at the end of the discussion to reflect this.

Significance

In this paper, the identification of BICD2 as a novel factor regulating centriole engagement is of significant importance. However, the mechanisms through which BICD2 controls its localization to the centrosome and regulates centriole engagement remain largely undefined. Further exploration of these mechanisms would likely enhance the value of the paper.

The findings are likely to be of great interest to researchers in the field of cell biology, particularly those focusing on centrosome biology.

The above feedback comes from a researcher specializing in centrosome studies.

__ __

Reviewer #2

Evidence, reproducibility and clarity

Montez-Ruiz and colleagues explore the role of a dynein adaptor BICD2 in the engagement of mother and daughter centrioles. Cells need to maintain centriole engagement in interphase to prevent centriole reduplication and in early mitosis to prevent the formation of aberrant mitosis spindles. The authors demonstrate that BICD2 is a centriolar protein that surrounds the mother centriole adjacent to the daughter centriole. It is removed from centrosomes in mitosis, which, in turn, is responsible for centriole disengagement. Further, they suggest that in BICD2 knock-out G2 and early mitotic cells, centrioles disengage prematurely. By conducting rescue experiments, the authors conclude that BICD2 regions CC2, CC3, and CC4, which are dynein-independent, are essential for their function at the centrosome. Finally, they show that the phosphorylation of S817 and S819 of BICD1 controls its centrosome localization.

Major comments:

1. __ Based on F1 and SF1, BICD2 is reduced from centrosomes already in early G2. So, it is hard to square how removing a factor that is not present at the centrosomes at the time of disengagement would dysregulate disengagement. The study at this stage does not explain how BICD2 contributes to centriole engagement only in mitosis, while it does not affect centrioles in S.__ We now present new data obtained using expansion microscopy (ExM) that, together with our super-resolution observations, clarifies this point. As shown in the new Figure 3 and Figure EV2 (and supported by Figures EV3 and EV4), although the total amount of BICD2 at centrosomes is significantly reduced from G2 to M, a pool of BICD2 persists at the mother centriole until late mitosis. Importantly, this pool tends to localize close to the daughter centriole. We note this in the text (“BICD2 remained visible in both diplosomes, associated with the SAS-6 foci (Figure 3, Figures EV2-3). Around anaphase, BICD2 was not detectable in some diplosomes, while others retained some protein (again, close to the SAS-6 foci, which at this point were disappearing from the centrioles as the result of the disassembly of the cartwheel).”). Supported by our BICD2 depletion experiments, we propose that this centriolar pool enables BICD2 to contribute to engagement until late mitosis, when the remaining protein at the centrosome is ultimately removed. We highlight this model in the Discussion section (“In mitosis, when the protein progressively disappears from the centrosomes, BICD2 remains functionally relevant -likely via the small pool that persists at the mother–daughter centriole interface.”).

Based on our new data demonstrating an interaction with CEP152, we propose that BICD2 forms an outer component of the centriolar torus. Centriole engagement is known to be maintained during S phase by the cartwheel (Huang F et al. 2022. Cartwheel disassembly regulated by CDK1-Cyclin B kinase allows human centriole disengagement and licensing. The Journal of Biological Chemistry 298:102658. DOI: https://doi.org/10.1016/j.jbc.2022.102658; Ito KK et al. 2025. Multimodal mechanisms of human centriole engagement and disengagement. The EMBO journal 44:1294–1321. DOI: https://doi.org/10.1038/s44318-024-00350-8), with the torus playing a role in cohesion later in the cell cycle. We note in our manuscript that this is consistent with our observations and supports a model in which BICD2 functions as part of the torus: “During S phase, mother–daughter centriole cohesion is maintained by the cartwheel (Huang et al, 2022; Ito et al, 2025) and, consistently, does not depend on BICD2.”

__ The interpretation that the longitudinal localization of the BICD2 signal coincides with SAS-6 and procentrioles requires further evidence. BICD2 seems largely localized to the other regions around the mother centriole, and in some examples, it does not colocalize with the site of the daughter centriole or SAS-6 (for instance: F2B second row; SF4B, second row; SF5, fourth row; SF6 upper row).__

We have added an ExM characterization of BICD2 centrosomal localization in the revised manuscript (Figures 2 and 3), that we think further clarify this point, showing that BICD2 longitudinally coincides with the torus and the daughter centriole. This is supported by new additional superresolution images (Figure 2, Figure EV2).

Note that ExM revealed an additional stable pool of BICD2 at the distal end of the centrioles that is not detected using standard methanol fixation combined with 3D-SIM. As discussed in the text this distal pool may reflect additional centriolar functions of BICD2.

__ BICD2 is important for centrosome-nucleus tethering during centrosome separation in G2, and its global removal likely affects the dynamics of the spindle assembly. Is G2 and mitotic progression affected in knockouts? Do the knockout cells show issues with chromosome alignment? Such analyses are critically missing from the manuscript.__

BICD2 knockout cells indeed show a slightly higher mitotic index than their wild type counterparts, and a higher frequency of lagging chromosomes in anaphase and telophase as well (new data, shown in Figure EV5C). We agree with the reviewer that these might result from the role of BICD2 tethering centrosomes to the nuclear envelope to facilitate their separation during the initial steps of spindle formation. We have added a sentence in the text noting this: “As expected from cells with supernumerary centrioles, BICD2-/- cells showed a slightly higher mitotic index and a higher frequency of lagging chromosomes in anaphase and telophase (Figure EV5C), although these mitotic defects might also be partially attributed to the role of BICD2 in centrosome separation (Splinter et al, 2010; Gallisà-Suñé et al, 2023).”

__ In general, SCLT experiments are ambiguous. Centriole disengagement spontaneously occurs during prolonged prometaphase induced by SCLT. Accordingly, F6D shows that many centriole pairs in the control sample are disengaged after 16h of SCLT treatment. Although the distance between centrioles in knockout cells is, on average, larger, without knowing how BICD2 perturbations affect the dynamics of the mitotic spindles and mitosis progression, SCLT experiments do not provide enough insight.__

After 16h of SCLT treatment, the authors regularly measure centriole distances in mitosis smaller than 500 nm in all samples. This suggests that the used method (which also needs to be described) cannot reliably assess centriole engagement status. Centrioles can be disengaged but adjacent. The authors reference Shukla et al. 2015 to compare the centriole-to-centriole distances here with those from that publication. However, in Shukla 2015, centriole-to-centriole distances increase from S to M. But here, in F6, the control centriole distances in S, G2, and early M are almost identical and less than 500 nm. This discrepancy needs to be addressed.

We thank the reviewer for these constructive comments. We appreciate the opportunity to further clarify our methodology and experimental rationale.

Validity and necessity of STLC treatment (Figures 4D and 7)

We fully agree with the reviewer that prolonged STLC treatment (16 hours) carries inherent limitations and should not serve as the sole experimental system for studying centriole engagement. As the reviewer notes, 16 hours of STLC treatment results in a baseline population of control cells displaying disengaged centrioles. This population likely represents cells that entered mitosis early during the treatment and remained arrested for the longest duration, or cells with inherently less robust engagement machinery.

However, we would like to highlight two key observations that validate STLC as a useful comparative tool in our study:

• The significative increase in the number of cells with higher intercentriolar distances that indicate disengagement in particular experimental conditions. We show that depletion of BICD2 consistently leads to a statistically significant increase in mean intercentriolar distances compared to controls under identical STLC conditions, indicating a distinct weakening of centriole engagement in a substantial number of cells (and thus suggesting that BICD2 is part of the engagement mechanism).

• Validation in unarrested cells: Crucially, this effect is not an artifact of mitotic arrest. Unarrested, normally cycling mitotic cells also display significantly increased intercentriolar distances in the absence of BICD2 (Figure 4C).

Following the initial characterization in Figure 4D, we restricted the use of STLC exclusively to experiments requiring cell transfection and recombinant protein expression (Figure 7). Human RPE-1 cells offer the key advantage of being an untransformed, p53-wild-type model. However, they also present technical challenges, including lower transfection efficiencies and sensitivity to experimental manipulation. Capturing a statistically robust sample of transfected, unarrested mitotic cells proved technically challenging. STLC treatment provided a necessary tool to enrich for mitotic cells while allowing clear observation of rescue effects.

We have explicitly clarified this technical rationale in the manuscript text:

"Although this treatment inherently increased mean intercentriolar distances, it nevertheless enabled clear observation of the effects of BICD2 ablation, while yielding a sufficient number of mitotic cells expressing the recombinant proteins."

Assessment of centriole engagement

We agree that centrioles can occasionally be disengaged while still remaining adjacent. To avoid oversimplifying the observed phenotypes, we chose to report raw intercentriolar distances rather than applying an arbitrary binary classification of "engaged" versus "disengaged." Furthermore, we do not rely solely on distance measurements to assess engagement status. We complemented these data by quantifying c-NAP1-positive centrioles in unarrested, cycling mitotic cells (Figure 4E, F). Because c-NAP1 loading marks centriole-to-centrosome conversion (and thus licensing), this functional readout independently confirms that BICD2 loss promotes premature centriole disengagement.

Intercentriolar distances across the cell cycle and cell-type variation

Regarding the comparison with Shukla et al. (2015), we note that their study was conducted in HeLa cells, whereas our primary model is RPE-1 (alongside U2OS cells). Variations in centriole engagement dynamics and distance kinetics can likely be attributed to intrinsic differences among these cell types:

RPE-1 cells: baseline intercentriolar distances in S-phase control RPE-1 cells (0.4–0.5 µm, measured using centrin) match those reported for HeLa cells in S-phase by Shukla et al. However, in RPE-1 control cells, these distances remain relatively constant from S phase through early M phase (Figure 4).

U2OS cells: U2OS cells exhibit a slight increase from 0.43±0.01 µm in G2 to 0.50±0.01 µm in M (Figure 5), illustrating that slight variations occur between cell lines.

Other studies similarly report persistent baseline distances around 0.5 µm through early cell cycle stages. For example, Yaguchi et al. (Yaguchi K et al. 2018. Uncoordinated centrosome cycle underlies the instability of non-diploid somatic cells in mammals. The Journal of Cell Biology 217:2463–2483. DOI: https://doi.org/10.1083/jcb.201701151) observed intercentriolar distances close to 0.5 µm in diploid HAP1 cells throughout mitosis and into early G1 phase, with substantial disengagement (>0.8 µm) occurring only well after cytokinesis onset.

To address this discrepancy, we have added the following sentence to the manuscript text: "Note that in wild-type S-phase RPE-1 cells, intercentriolar distances measured using centrin as a marker were similar to those described in S-phase HeLa cells (Shukla et al., 2015), namely 0.4–0.5 µm; however, in contrast to HeLa cells, these distances remained fairly constant from S to early M phase in RPE-1 cells." We have also updated the Materials and Methods section to provide a precise description of how intercentriolar distances were measured: “Intercentriolar distances were assessed as the distance between centrin foci of the same diplosome in maximum projections of z-stacks”

__ The authors suggest that BICD2's functions at the centrosome are independent of its dynein functions. They show that GFP-BICD2 1-820 DD rescues centriole engagement among several other mutants. However, it is still possible that the expression of the mutants affects some yet uncovered BICD2 function outside of centrosomes. At least, T821A and S823A should be mutated to Ala. From what I gathered, such mutant should remain associated with mitotic centrosomes. The authors should analyze whether mitotic progression remains unperturbed, and centriole engagement status should be analyzed without SCLT treatment in G2, M, and in ensuing G1.__

We agree with the reviewer that phosphonull mutants should be added to these experiments. In fact, and as mentioned in the responses to Reviewer 1, we have already performed experiments with the phosphonull mutants, observing that they are more retained at centrosomes than the phosphomimetic counterparts. We would be happy to share these results with the reviewers upon request if helpful. Nevertheless, and as mentioned above, we have decided to remove the preliminary data regarding BICD2 phosphorylation from the manuscript data in order to present a separate and more comprehensive study on BICD2 phosphorylation in the near future.

Significance

The question explored is relevant to the centrosome field and beyond since the processes leading to premature centriole disengagement and amplification are not fully understood. The study provides some novel insights. However, at the current stage, the study is preliminary. Additional experiments would be needed to strengthen the conclusion that BICD2 directly regulates centriole disengagement.

My expertise is in centriole and centrosome assembly and the mechanisms that regulate centrosome homeostasis in human cells.

__ __

__Reviewer #3 __

Evidence, reproducibility and clarity (Required):

Centrosome duplication is tightly control during cell cycle to prevent loss or amplification of centrosome numbers, which are detrimental for cell proliferation. In preparation for centriole duplication in S-phase, mother and daughter centrioles disengaged late mitosis, a process that functions as a licensing factor for duplication. While several mechanism have been proposed to be important for centriole disengagement, differences between systems and organisms exist, suggesting alternative pathways may play a role.

In this manuscript, Montes-Ruiz and colleagues investigate the role of the dynein adaptor protein BICD2 during centriole disengagement. They found that BICD2 localises to the centrioles, with a peak in S-Phase. Super resolution microscopy suggests that BICD2 localises to the mother centrioles and is mostly absent in mitosis cells after anaphase, when centrioles are disengaging. KO of BICD2 in REP-1 cells does not some t have strong phenotypes, but the authors found that centriole separation is increased, suggesting a role in centriole cohesion. While there is limited mechanist insight about the regulation of BICD2 and its function at the centrosomes, the data presented suggests a role for BICD2 in centriole cohesion that is independent of dynein interaction. There are however several issues with data presentation, image analyses and data interpretation the authors could improve.

Major comments

- On page 5, the authors state that figure 1 and supplementary figure1 data strongly suggest that BICD2 associates with mother centrioles and not the PCM. This is not very clear from the images on these figures. In fact, PCM is often associated with mother centriole as well, thus I am not sure they can make these conclusions based on the data presented in these 2 figures. Also, the fact that PCM is more abundant in G2/M, when BICD2 is not, does not mean it does not localize to the PCM. Higher resolution of expansion will be needed.

The data presented in supplementary figure 3 does not help the conclusion above as it seems form the images that there is co-localization between BICD2 and pericentrin. It is impossible to conclude also that there is co-localization with the satellite marker PCM-1. In fact, they seem to have no overlap from the images provided. Higher resolution of expansion will be needed.

Following the reviewer’s suggestion we embarked in a full characterization of BICD2 localization using expansion microscopy (ExM). We think that our new data further clarifies this together with new superresolution data.

Additionally we now have a figure (Supplementary Figure S4) addressing the relation between pericentrin and the localization of BICD2. We show that pericentrin downregulation does not affect centrosomal BICD2 levels. And that both proteins do not colocalize as observed using 3D-SIM.

Also regarding pericentrin, the revised version of the manuscript now includes a figure that functionally compares the results of its depletion to those of BICD2 (Figure 5).

We also provide data showing that BICD2 localization does not significatively change upon PCM-1 depletion (Supplementary Figure S4C). As we mention in the text, previous reports have suggested that BICD2 is indeed in the satellites (Quarantotti V et al. 2019. Centriolar satellites are acentriolar assemblies of centrosomal proteins. The EMBO Journal e101082. DOI: https://doi.org/10.15252/embj.2018101082) . But we only observed clear colocalization of PCM-1 with C-terminal fragments of BICD2. Thus, while GFP-BICD2 488–820 strongly colocalizes with satellites, endogenous BICD2 and full-length GFP-BICD2 do not (Figure 6D). We ignore the reason for this, but the data suggests that satellite localization is regulated (possibly via intramolecular autoinhibition) to mediate BICD2 transport or a distinct satellite-specific function of this protein. To address this we have added a sentence to the text that now reads: “The presence of BICD2 at the centriolar satellites has been suggested previously (Quarantotti et al, 2019); we ignore the reason why in the conditions used in this study only C-terminal fragments of BICD2 (but not the full-length protein, see Supplementary Figure S4) colocalize with satellites. This suggests that BICD2 satellite localization is regulated (possibly via intramolecular autoinhibition) to mediate BICD2 transport or a distinct satellite-specific function of this protein.”.

- In figure 2, to confirm localization to the mother centrioles, could the authors use a mother centriole marker? Such as a distal appendage protein of ninein? CEP152 localizes to both centrioles in the images provided.

I was surprised that BICD2 localizes to both distal appendages and linker? These are not close to each other. Can the authors comment on this? In supplementary figure 4C orthogonal view it seems like BICD2 is in between distal appendages and linker?

We believe that the new ExM data (Figures 2 and 3) directly address the reviewer's concerns.

Regarding the original supplementary figure S4C, indeed in the orthogonal projections of 3D-SIM images the signal corresponding to BICD2 was observed between distal appendages and linker and was quite broadly distributed. We recognize that this could lead to confusion. We have now removed part of this figure (original Figures 4B and 4C) from the manuscript, as we think that the data is made redundant with our new ExM data. Our new data, with a much higher resolution shows that BICD2 localization corresponds to that of the proximal torus (see new Figures 2D and 2E, and Figure 3). Note that in our new ExM images we use a daughter centriole marker (SAS-6) that (in addition to CEP152) we think helps confirm that BICD2 localizes around the mother centriole.

-The IF data suggests that BICD2 localization to the centrosome is dynamically regulated during cell cycle. Did the authors consider that this protein could be degraded? Is it a matter of recruitment or total protein levels?

We agree that protein degradation has to be considered when analyzing cell cycle-dependent localization. However, our data suggest that the dynamic behaviour of BICD2 at the centrosomes does not reflect changes in its total protein amount. We have previously shown that total BICD2 levels are not reduced in mitosis, as assessed by western blot (Gallisà-Suñé N et al. 2023. BICD2 phosphorylation regulates dynein function and centrosome separation in G2 and M. Nature Communications 14:2434. DOI: https://doi.org/10.1038/s41467-023-38116-1). To make this clear in the current manuscript, we have additionally added Figure EV1B depicting BICD2 levels in S, G2 and M phase, and the following note to the text : “Total levels of BICD2 remained constant during the different phases of the cell cycle (Figure EV1B and (Gallisà-Suñé et al, 2023))“.

- The authors propose that the dynamic localization of BICD2 is associated with licensing. However, it is rather surprising that the phenotype of centriole separation they describe is only observe in mitosis when BICD2 in knockdown and not in S-phase when the levels of BICD2 are higher? If the role of BICD2 is to prevent premature centiole disengagement, shouldn't that be observed in S-phase as well? Why only in mitosis when in control cells BICD2 levels are already very low?

Recent data supports the notion that in S phase centriole cohesion is maintained by the cartwheel (Huang F et al. 2022. Cartwheel disassembly regulated by CDK1-Cyclin B kinase allows human centriole disengagement and licensing. The Journal of Biological Chemistry 298:102658. DOI: https://doi.org/10.1016/j.jbc.2022.102658; Ito et al. 2025. Multimodal mechanisms of human centriole engagement and disengagement. The EMBO journal 44:1294–1321. DOI: https://doi.org/10.1038/s44318-024-00350-8). Our data, including the new results showing that BICD2 interacts with CEP152, suggests that BICD2 is a dynamic part of the mother centriole torus, a structure that does not seem to be implicated in maintaining cohesion in S. We now note this in the manuscript’s text: “During S phase, mother-daughter centriole cohesion is maintained by the cartwheel (Huang et al, 2022; Ito et al, 2025), and, consistently, does not depend on BICD2.”. Of note, after Ito etl al. BICD2 (and the torus) may have a role in late S if the cartwheel is compromised, something that could be tested in future studies by downregulating cartwheel components and BICD2 simultaneously.

- The images of C-Nap1 localization in figure 6E are not very convincing to illustrate the pint the authors are making in the main text (additional C-Nap1 foci are visible in the KO cells)

We would like to note that visualizing C-NAP1 in mitosis is technically challenging, as a significant pool of the protein is displaced from the centrioles after phosphorylation in G2. However, the protein has been widely used as a marker of centriole disengagement (e.g. in the seminal Tsou M-FB et al. 2006. Mechanism limiting centrosome duplication to once per cell cycle. Nature 442:947–951. DOI: https://doi.org/10.1038/nature04985). We therefore consider it a valuable tool to support our conclusions regarding centriole engagement. Regarding extra c-NAP1 foci in BICD2 knockout cells, these may reflect additional centrioles that appear in these cells, as a result of abnormal disengagement and early licensing. To have this into account our data quantifies both c-NAP-1 positive centrioles (increased in KO cells, Figure 4E) and number of c-NAP-1 positive centrioles /total centriole number (with an increase in the abnormal >2:4 configuration in BICD2 KO cells, Figure 4F).

- The authors propose that PLK1 and CDK1 phosphorylation sites regulate the association of BICD2 with the centrioles. Could this be tested with a PLK1 inhibitor?

As noted above, we have removed the phosphorylation data from the manuscript, as we aim to report these findings in a dedicated upcoming study. Nevertheless, to address the reviewer's query, we now consider BICD2 to be predominantly regulated by CDK1, supported by data using BI 2536 showing that BICD2 centrosomal levels are unaffected by PLK1 inhibition. In contrast, CDK1 inhibition slightly reduces these levels, though this effect does not reach statistical significance under the tested conditions. We would be glad to share these additional results with the reviewers upon request.

- On page 13, the authors state that their results do not agree with previous literature showing that pericentrin cleavage can result in disengagement. However, it was unclear from this manuscript what is the evidence to demonstrate that this is the case? The data presented in figure 5B for example only demonstrates that pericentrin levels do not change in the absence of BICD2 in what looks like S-phase cells. Did the authors look at pericentrin levels when they observe centriole disengagement in the ko cells? In G2 or early M-phase?

We recognize that pericentrin is widely considered a crucial factor in centriole engagement, and have added new data in the manuscript studying the relationship between it and BICD2 (the partially new Supplementary Figure S4), and their relative importances for engagement both in G2 and M (the new Figure 5). Our data suggests that both proteins act independently in a partially redundant manner, BICD2 as part of the torus (key for engagement in G2) and pericentrin of the PCM (more important in M).

We have updated the Discussion to present this and our view on pericentrin importance for engagement more clearly, specially our concerns that its importance may have been overestimated. Specifically we write that “BICD2 depletion reduces its centrosomal levels to a degree that mirrors those naturally observed during late M and early G1 in unperturbed cells. In contrast, experimental depletion of pericentrin reduces its levels far below physiological baselines across any phase of the cell cycle. This severe reduction produces marked centriole separation in mitosis that is likely amplified by spindle-derived forces. Consequently, the individual contribution of pericentrin to regulating physiological centriole cohesion may be somewhat overestimated under standard experimental knockdowns, and this regulation may rely more heavily on torus components, such as BICD2, than previously appreciated.”

Regarding the phases of the cell cycle in which we quantify pericentrin levels in the original Figure 5B (now Figure EV5B), we realize that the figure could lead to confusion as it was, as they were measured in M (when its amount is maximal, as specified in the figure legend) but the figure did not show examples in this cell cycle phase. We have added new examples of mitotic cells to the figure, and modified the figure labels and wording of the figure legend to clarify this.

Minor comments

- A more general reference (review) missing in the second paragraph of the introduction that describe the centrosomes.

We have added a recent general reference when introducing centrioles (Gönczy P. 2025. Critical constituents and assembly principles of centriole biogenesis in human cells. Nature Reviews Molecular Cell Biology 1–18. DOI: https://doi.org/10.1038/s41580-025-00921-5). Later in the paragraph, when centriole duplication is introduced, we now use this reference plus the also recent Fernandes-Mariano C et al. 2025. Centrosome biogenesis and maintenance in homeostasis and disease. Current Opinion in Cell Biology 94:102485. DOI: https://doi.org/10.1016/j.ceb.2025.102485.

- Some figures are not well organized, difficult to see which panel they correspond to? The authors could consider labelling panels better to make this clear. For example, figure 4 and 6 could benefit from additional panel labels.

We have added additional panel labels to Figure 4 (now Figure 6) and Figure 6 (now Figure 4), that we have also slightly reorganized with the aim of making it clearer).

- On page 7, what the authors mean by: "... we ignore the reason why in the conditions used in this study only C-terminal fragments of BICD2 but not the fill-length protein co-localize with these pericentriolar structures"?

By "pericentriolar structures” we were referring to the centriolar satellites. We realize that that was not clear and updated the wording of the sentence that now reads “we ignore the reason why in the conditions used in this study only C-terminal fragments of BICD2 (but not the full-length protein, see Supplementary Figure S4) colocalize with satellites.”. We subsequently propose a possible a possible explanation for this: "This suggests that BICD2 satellite localization is regulated (possibly via intramolecular autoinhibition) to mediate BICD2 transport or a distinct satellite-specific function of this protein.".

Reviewer #3 (Significance (Required)):

In general this work has limited mechanistic insight and BICD2 localization to the centrosomes was known. However, the authors do go into more detail description of the centriole localization of BICD2 . In addition, their established KO cell lines provide some insights into the role of BICD2 in centriole disengagement, which is of interest to the field. But the limited scope of the conclusions does not advance the field significantly as it is.

this work will interest a specialized audience.

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Referee #3

Evidence, reproducibility and clarity

Centrosome duplication is tightly control during cell cycle to prevent loss or amplification of centrosome numbers, which are detrimental for cell proliferation. In preparation for centriole duplication in S-phase, mother and daughter centrioles disengaged late mitosis, a process that functions as a licensing factor for duplication. While several mechanism have been proposed to be important for centriole disengagement, differences between systems and organisms exist, suggesting alternative pathways may play a role. In this manuscript, Montes-Ruiz and colleagues investigate the role of the dynein adaptor protein BICD2 during centriole disengagement. They found that BICD2 localises to the centrioles, with a peak in S-Phase. Super resolution microscopy suggests that BICD2 localises to the mother centrioles and is mostly absent in mitosis cells after anaphase, when centrioles are disengaging. KO of BICD2 in REP-1 cells does not some t have strong phenotypes, but the authors found that centriole separation is increased, suggesting a role in centriole cohesion. While there is limited mechanist insight about the regulation of BICD2 and its function at the centrosomes, the data presented suggests a role for BICD2 in centriole cohesion that is independent of dynein interaction. There are however several issues with data presentation, image analyses and data interpretation the authors could improve.

Major comments

On page 5, the authors state that figure 1 and supplementary figure1 data strongly suggest that BICD2 associates with mother centrioles and not the PCM. This is not very clear from the images on these figures. In fact, PCM is often associated with mother centriole as well, thus I am not sure they can make these conclusions based on the data presented in these 2 figures. Also, the fact that PCM is more abundant in G2/M, when BICD2 is not, does not mean it does not localize to the PCM. Higher resolution of expansion will be needed. The data presented in supplementary figure 3 does not help the conclusion above as it seems form the images that there is co-localization between BICD2 and pericentrin. It is impossible to conclude also that there is co-localization with the satellite marker PCM-1. In fact, they seem to have no overlap from the images provided. Higher resolution of expansion will be needed. In figure 2, to confirm localization to the mother centrioles, could the authors use a mother centriole marker? Such as a distal appendage protein of ninein? CEP152 localizes to both centrioles in the images provided. I was surprised that BICD2 localizes to both distal appendages and linker? These are not close to each other. Can the authors comment on this? In supplementary figure 4C orthogonal view it seems like BICD2 is in between distal appendages and linker? The IF data suggests that BICD2 localization to the centrosome is dynamically regulated during cell cycle. Did the authors consider that this protein could be degraded? Is it a matter of recruitment or total protein levels? The authors propose that the dynamic localization of BICD2 is associated with licensing. However, it is rather surprising that the phenotype of centriole separation they describe is only observe in mitosis when BICD2 in knockdown and not in S-phase when the levels of BICD2 are higher? If the role of BICD2 is to prevent premature centiole disengagement, shouldn't that be observed in S-phase as well? Why only in mitosis when in control cells BICD2 levels are already very low? The images of C-Nap1 localization in figure 6E are not very convincing to illustrate the pint the authors are making in the main text (additional C-Nap1 foci are visible in the KO cells) The authors propose that PLK1 and CDK1 phosphorylation sites regulate the association of BICD2 with the centrioles. Could this be tested with a PLK1 inhibitor? On page 13, the authors state that their results do not agree with previous literature showing that pericentrin cleavage can result in disengagement. However, it was unclear from this manuscript what is the evidence to demonstrate that this is the case? The data presented in figure 5B for example only demonstrates that pericentrin levels do not change in the absence of BICD2 in what looks like S-phase cells. Did the authors look at pericentrin levels when they observe centriole disengagement in the ko cells? In G2 or early M-phase?

Minor comments

A more general reference (review) missing in the second paragraph of the introduction that describe the centrosomes. Some figures are not well organized, difficult to see which panel they correspond to? The authors could consider labelling panels better to make this clear. For example, figure 4 and 6 could benefit from additional panel labels. On page 7, what the authors mean by: "... we ignore the reason why in the conditions used in this study only C-terminal fragments of BICD2 but not the fill-length protein co-localize with these pericentriolar structures"?

Significance

In general this work has limited mechanistic insight and BICD2 localization to the centrosomes was known. However, the authors do go into more detail description of the centriole localization of BICD2 . In addition, their established KO cell lines provide some insights into the role of BICD2 in centriole disengagement, which is of interest to the field. But the limited scope of the conclusions does not advance the field significantly as it is.

this work will interest a specialized audience.

Note: This preprint has been reviewed by subject experts for Review Commons. Content has not been altered except for formatting.

Learn more at Review Commons

Referee #2

Evidence, reproducibility and clarity

Montez-Ruiz and colleagues explore the role of a dynein adaptor BICD2 in the engagement of mother and daughter centrioles. Cells need to maintain centriole engagement in interphase to prevent centriole reduplication and in early mitosis to prevent the formation of aberrant mitosis spindles. The authors demonstrate that BICD2 is a centriolar protein that surrounds the mother centriole adjacent to the daughter centriole. It is removed from centrosomes in mitosis, which, in turn, is responsible for centriole disengagement. Further, they suggest that in BICD2 knock-out G2 and early mitotic cells, centrioles disengage prematurely. By conducting rescue experiments, the authors conclude that BICD2 regions CC2, CC3, and CC4, which are dynein-independent, are essential for their function at the centrosome. Finally, they show that the phosphorylation of S817 and S819 of BICD1 controls its centrosome localization.

Major comments:

1. Based on F1 and SF1, BICD2 is reduced from centrosomes already in early G2. So, it is hard to square how removing a factor that is not present at the centrosomes at the time of disengagement would dysregulate disengagement. The study at this stage does not explain how BICD2 contributes to centriole engagement only in mitosis, while it does not affect centrioles in S.
2. The interpretation that the longitudinal localization of the BICD2 signal coincides with SAS-6 and procentrioles requires further evidence. BICD2 seems largely localized to the other regions around the mother centriole, and in some examples, it does not colocalize with the site of the daughter centriole or SAS-6 (for instance: F2B second row; SF4B, second row; SF5, fourth row; SF6 upper row).
3. BICD2 is important for centrosome-nucleus tethering during centrosome separation in G2, and its global removal likely affects the dynamics of the spindle assembly. Is G2 and mitotic progression affected in knockouts? Do the knockout cells show issues with chromosome alignment? Such analyses are critically missing from the manuscript.
4. In general, SCLT experiments are ambiguous. Centriole disengagement spontaneously occurs during prolonged prometaphase induced by SCLT. Accordingly, F6D shows that many centriole pairs in the control sample are disengaged after 16h of SCLT treatment. Although the distance between centrioles in knockout cells is, on average, larger, without knowing how BICD2 perturbations affect the dynamics of the mitotic spindles and mitosis progression, SCLT experiments do not provide enough insight. After 16h of SCLT treatment, the authors regularly measure centriole distances in mitosis smaller than 500 nm in all samples. This suggests that the used method (which also needs to be described) cannot reliably assess centriole engagement status. Centrioles can be disengaged but adjacent. The authors reference Shukla et al. 2015 to compare the centriole-to-centriole distances here with those from that publication. However, in Shukla 2015, centriole-to-centriole distances increase from S to M. But here, in F6, the control centriole distances in S, G2, and early M are almost identical and less than 500 nm. This discrepancy needs to be addressed.
5. The authors suggest that BICD2's functions at the centrosome are independent of its dynein functions. They show that GFP-BICD2 1-820 DD rescues centriole engagement among several other mutants. However, it is still possible that the expression of the mutants affects some yet uncovered BICD2 function outside of centrosomes. At least, T821A and S823A should be mutated to Ala. From what I gathered, such mutant should remain associated with mitotic centrosomes. The authors should analyze whether mitotic progression remains unperturbed, and centriole engagement status should be analyzed without SCLT treatment in G2, M, and in ensuing G1.

Significance

The question explored is relevant to the centrosome field and beyond since the processes leading to premature centriole disengagement and amplification are not fully understood. The study provides some novel insights. However, at the current stage, the study is preliminary. Additional experiments would be needed to strengthen the conclusion that BICD2 directly regulates centriole disengagement.

My expertise is in centriole and centrosome assembly and the mechanisms that regulate centrosome homeostasis in human cells.

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Referee #1

Evidence, reproducibility and clarity

This study demonstrates that BICD2, previously known as an adaptor protein for dynein, is involved in regulating centriole engagement during mitosis. First, using different antibodies, it was shown that BICD2 localizes near the mother centriole, as observed by super-resolution microscopy. During G1 and S phases, BICD2 localizes slightly outside the Cep152 ring, while in G2 to mitosis, it localizes near the cartwheel component SAS-6. Moreover, analysis of deletion mutants revealed that BICD2 localizes to the centrosome in a CC domain-dependent manner at the C-terminal end. The localization pattern resembling a ring in the cytoplasm was also observed through the CC3 domain. Next, BICD2 knockout (KO) cells were generated to investigate centriole dynamics. In BICD2 KO cells, the distance between the mother and daughter centrioles was observed to increase from G2 to mitosis compared to controls. Along with this, early centriole disengagement and centriole amplification phenotypes were observed. The increased distance phenotype between centrioles was rescued in BICD2 wild-type (WT) and mutant forms lacking the CC1 domain at the N-terminus, suggesting that this function of BICD2 is independent of dynein. Additionally, BICD2 mutants mimicking phosphorylation at the C-terminus showed reduced centrosome localization and were unable to rescue the phenotypes seen in BICD2 KO cells. While the study clearly demonstrates BICD2's contribution to centriole engagement, the underlying mechanisms of how BICD2 is involved in centrosome localization and centriole engagement remain unclear. As it is anticipated that the function of BICD2 is independent of dynein, further exploration of this unknown mechanism would enhance the value of the paper. Below are the concerns that should be addressed, including new experiments.

Main Points:

1. Fig. 1-3: Regarding the localization of BICD2 to centrioles, during the G1-S phase, its localization appears to overlap with PCM. Experimental investigation should be performed to examine whether BICD2's centrosomal localization is influenced by knockdown of PCM components like PCNT, Cep192, or Cep152.
2. Fig. 7: The experiments using BICD2 mutants suggest that the function of BICD2 here is independent of dynein. To further investigate whether BICD2's role in centriole engagement is independent of dynein, experiments should be conducted to examine the effect of dynein knockdown on BICD2 localization to the centrosome and centriole engagement.
3. Fig. 4: The CC4 domain at the C-terminus of BICD2 is important for its centrosomal localization, but identifying the binder/recruiter responsible for BICD2's centrosome localization would be desirable.
4. Fig. 7: Rescue experiments using BICD2 mutants suggest that BICD2's functional domains are critical. Further experiments by creating mutants missing parts of CC2 or CC3 could identify functionally important domains of BICD2 by observing any loss-of-function phenotypes at the centrosome.
5. Fig. 6: Regarding the BICD2 KO cell phenotype, is there experimental evidence showing an increase in centriole number during mitosis? For instance, while no abnormality in centriole number may occur during G2, a trend of increase in mitosis should be experimentally demonstrated. Also, how should the slight differences in phenotypes between Ndelta4 and Ndelta5 BICD2 KO cells be interpreted?
6. Fig. 8: Regarding the phosphorylation of BICD2 at the C-terminus: The phenotypes of mutants where these two phosphorylation sites are changed to alanine should be experimentally observed. It is expected that the removal of BICD2 from the centrosome during mitosis could be rescued. Additionally, the effect of PLK1 or CDK1 inhibitors on the removal of BICD2 from the centrosome should be investigated.

Minor Points:

1. Fig. 1-3: During G1 and S phases, BICD2 localizes near the mother centriole, and from G2 onward, it colocalizes with SAS-6. How can this be explained?
2. Fig. 4: The GFP-BICD2 488-820 fragment forms cytoplasmic rings, which is interesting. This domain contains the CC4 domain, so it can localize near the centriole, but why does it not form a perfect ring there? Also, which other centriole/centrosome markers were used for colocalization studies? Does knockdown of PCM1 affect BICD2's centrosomal localization?
3. Fig. 4A: What are the aggregates observed in the cytoplasm under the GFP-BICD2 + ice condition? Also, does the 1-575 mutant fail to localize to the centrosome upon ice treatment?
4. Can similar phenotypes be observed in other cell types when BICD2 is knocked down? This should be experimentally validated.
5. Are there previous studies suggesting that this function of BICD2 is evolutionarily conserved? This should be addressed.

Significance

In this paper, the identification of BICD2 as a novel factor regulating centriole engagement is of significant importance. However, the mechanisms through which BICD2 controls its localization to the centrosome and regulates centriole engagement remain largely undefined. Further exploration of these mechanisms would likely enhance the value of the paper.

The findings are likely to be of great interest to researchers in the field of cell biology, particularly those focusing on centrosome biology.

The above feedback comes from a researcher specializing in centrosome studies.

O juiz das garantias será INVESTIDO, e não designado.

Os autos que compõem as matérias de competência do juiz das garantias serão remetidos ao juiz da instrução e julgamento

OFERECIMENTO da denúncia ou queixa

OFERECIDA a denúncia ou queixa, as questões pendentes serão decididas pelo juiz da instrução e julgamento

O STF decidiu, por unanimidade, atribuir interpretação conforme ao § 2º do art. 3º-B do CPP, incluído pela Lei nº 13.964/2019, para assentar que: a) o juiz pode decidir de forma fundamentada, reconhecendo a necessidade de novas prorrogações do inquérito, diante de elementos concretos e da complexidade da investigação; e b) a inobservância do prazo previsto em lei não implica a revogação automática da prisão preventiva, devendo o juízo competente ser instado a avaliar os motivos que a ensejaram, nos termos da ADI nº 6.581.

O STF deciciu, por maioria, declarar a inconstitucionalidade do inciso XIV do art. 3º-B do CPP, incluído pela Lei nº 13.964/2019, e atribuir interpretação conforme para assentar que a competência do juiz das garantias cessa com o oferecimento da denúncia, vencido o Ministro Edson Fachin.

O STF decidiu, por unanimidade, atribuir interpretação conforme ao inciso VII do art. 3º-B do CPP, incluído pela Lei nº 13.964/2019, para estabelecer que o juiz pode deixar de realizar a audiência quando houver risco para o processo, ou diferi-la em caso de necessidade.

Hay que construir una seccion explicando el Arrenius, en un capitulo previo

agregar los 3 casos de estudio

IBM Selectric Service Training (1978, Restored)<br /> by [[Thickboy]] on YouTube<br /> accessed on 2026-07-31T10:13:34

I just finished a project and I think a lot of you would like it. I found the old Selectric training videos but the narration is very outdated, slow paced and the slides were all burned red and miscolored.

With the help of AI I restored the photos and created a new narrator. The script has been modernized to sound more palatable to our ear. "Stop the reel" updated to "pause the video", for example and is now narrated at a natural pace in a woman's voice.

These courses, for me at least, are way easier to sit through and absorb. There maybe be some coloration errors or glitches in some photos, but for the most part they came out pretty good. Take a look and let me know what you think.

I'm also putting together some review videos with quiz questions for each section that I will release soon.

IBM Selectric Service Training (1978, Restored)<br /> via u/Mike3521 at https://www.reddit.com/r/typewriters/comments/1vbcsob/just_finished_restoring_the_old_selectric/

optional

a sustainable end-of-life practice

Day 4 launch day? Strategy note

building sustainable businesses around their end-of-life work, and how you can too.

learn practical strategies that lead to

If video ends up being removed, keep this!

missed link for VAU

we need to add comment that this method not supported for vau backups

we also have available option "--log-path" which defines where veeamsupporttool will save its execution log

Get started with your first little farm project right away!

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again, make this folded by default

this is minor -- make this box folded by default

this clump is hard to read here

So, from my understanding, the idea of 'filial piety' is sort of a cultural or religious practice where there is a certain expectation held over the children. Who cares for who depends on who needs the care, and it's assumed that the adult children will reciprocate (balanced!) the care to their parents, as a repayment for when they were younger.

My first question, which I'm hoping will be answered later in this article - are families hiring migrant workers, or are they the only ones willing to take on the care of Shanghai elders because of other reasons? Also, this tells me that the people likely being researched for this article are Chinese elders and migrant caregivers.

I think the author is trying to say here that building a rapport with non-familial relatives can, over time, create a bond just as strong as one between blood relatives.

Deeper (platonic or familial) relationships form due to more intimate nature of providing care to another person, at least when the caregiver has some empathy.

research takes place in Shanghai, China, one of the country's largest & most urbanized cities.

I think what this line is trying to say is that this article will have some first person POV's from the migrant care workers, and how they have formed kinship, or a close relationship, almost familial, with the elders they care for due to how closely they work with them.

This line reminded me of the two years I spent volunteering in a nursing home. Getting to know the elders of my community was an experience I wouldn't trade for anything. They're so full of knowledge and inspiration.

Note that this enzyme is also critical to the buffering of our blood's pH and thus preventing alkalosis or acidosis most of the time

For example it can be like the CFC shown in the previous example in which the Cl was provided in gaseuous form similar to ozone which was also a gas

fairly dangerous thing to do. this would also be conversion: from belief in Judaism to belief in enlightenment. and there was a hardy synthesis of those two beliefs, but it did not involve professing the dissolution of Christianity. more like the aesthetic adherence to Christianity.

what is social power?

Marx seems not to go into detail on what this final human emancipation is

the Jewish state less invested in having a leader with a divine mandate and more interested in possession of the land as primary link to heaven. extremely convenient, then, to have mentorship in colonization during the mandate years

still can't dump the god-king

applying this to Zionist beliefs: the Jew must be elevated to the status of citizen, the Jewish faith must be subsumed into a state, and that state must retain a majority Jewish population in order to say there is no difference between Judaism and status as a citizen of Israel

curious about this: does it create a situation where state leaders are free from a power struggle with church leaders?

thinking also about the cult of the state that has emerged in the US: without competition, the state can offer something in which to belief that both A: verifiably exists and B: can be influenced and paid tribute to through taxes, voting, other religious rituals.

grey zone in column of cumul à la fin du trimestre pour chaque année

add date when we receive it

link

link

link

full bullet point? see Q3 2025-26

add Pdf version when we have it final

make it easier/more prominent to 'clear all filters'

It was God's will to make Texas a part of America -- powers trying to counter American expansion, namely Britain and France, are against God's will and are hostile

Is this line necessary to this item? Can this be removed? Thanks.

add grey background align with column year to date at quarter end for both year

Replace by : Significant changes in relation to operations, personnel and programs

add date when received

add date when received

Link

link

add link

the bullet point need to be full? they were not in Q3 of last quarter

can we just put a summary data table like in the past exercice? or it's mandatory to put a description overview?

Add the PDF link when we send it

I researched this case. It does not appear so clear of a biased call (and the polls were internal to the campaign)

This work is now published as follows: Journal Nature Neuroscience DOI 10.1038/s41593-026-02383-0

Alucinaciones del 17%, incluso con Grounding (¿jurisprudencia EEUU?)

نیازمند اتصال به اینترنت تبدیل PDF به صورت صفحه به صفحه قابل استفاده در وب‌سایت و به صورت وب‌سرویس

IndyLab telegram channel 8 May 2026

screenshot

you should never believe that something is impossible to to do

in America The Impossible is what we do best

watch clip

see transcipt

singularity

impossible dream

DOI: 10.3390/ph19071080

Resource: (BCRC Cat# 60343, RRID:CVCL_0547)

Curator: @evieth

SciCrunch record: RRID:CVCL_0547

What is this?

DOI: 10.3390/ph19071080

Resource: (ATCC Cat# CCL-62, RRID:CVCL_1905)

Curator: @evieth

SciCrunch record: RRID:CVCL_1905

What is this?

DOI: 10.3390/ph19071080

Resource: (Ximbio Cat# 152707, RRID:CVCL_1941)

Curator: @evieth

SciCrunch record: RRID:CVCL_1941

What is this?

DOI: 10.3390/ph19071080

Resource: (CCLV Cat# CCLV-RIE 1035, RRID:CVCL_0023)

Curator: @evieth

SciCrunch record: RRID:CVCL_0023

What is this?

DOI: 10.3390/ph19071080

Resource: (NCI-DTP Cat# MCF7, RRID:CVCL_0031)

Curator: @evieth

SciCrunch record: RRID:CVCL_0031

What is this?

DOI: 10.3390/ph19071080

Resource: (CLS Cat# 300141/p1546_Calu-1, RRID:CVCL_0608)

Curator: @evieth

SciCrunch record: RRID:CVCL_0608

What is this?

DOI: 10.3390/ph19071080

Resource: (ICLC Cat# HTL95023, RRID:CVCL_0030)

Curator: @evieth

SciCrunch record: RRID:CVCL_0030

What is this?

DOI: 10.3390/ph19071080

Resource: (ECACC Cat# 89050205, RRID:CVCL_0548)

Curator: @evieth

SciCrunch record: RRID:CVCL_0548

What is this?

DOI: 10.3390/ph19071080

Resource: (RRID:CVCL_A8EZ)

Curator: @evieth

SciCrunch record: RRID:CVCL_A8EZ

What is this?

DOI: 10.3390/ph19071080

Resource: (BCRJ Cat# 0382, RRID:CVCL_0543)

Curator: @evieth

SciCrunch record: RRID:CVCL_0543

What is this?

DOI: 10.3390/ph19071080

Resource: (RRID:CVCL_0134)

Curator: @scibot

SciCrunch record: RRID:CVCL_0134

What is this?

DOI: 10.3390/ph19071080

Resource: (BCRJ Cat# 0395, RRID:CVCL_0168)

Curator: @scibot

SciCrunch record: RRID:CVCL_0168

What is this?

DOI: 10.3390/ph19071048

Resource: (RRID:CVCL_0063)

Curator: @evieth

SciCrunch record: RRID:CVCL_0063

What is this?

DOI: 10.3390/ph19071048

Resource: (NCI-DTP Cat# MCF7, RRID:CVCL_0031)

Curator: @scibot

SciCrunch record: RRID:CVCL_0031

What is this?

DOI: 10.1158/1078-0432.CCR-25-4596

Resource: Pindel (RRID:SCR_000560)

Curator: @evieth

SciCrunch record: RRID:SCR_000560

What is this?

DOI: 10.1126/sciadv.aef6777

Resource: RRID:IMSR_JAX:030170

Curator: @evieth

SciCrunch record: RRID:IMSR_JAX:030170

What is this?

DOI: 10.1126/sciadv.aef6777

Resource: (IMSR Cat# JAX_005628,RRID:IMSR_JAX:005628)

Curator: @evieth

SciCrunch record: RRID:IMSR_JAX:005628

What is this?

DOI: 10.1038/s41590-026-02544-0

Resource: (BioLegend Cat# 310914, RRID:AB_314849)

Curator: @evieth

SciCrunch record: RRID:AB_314849

What is this?

DOI: 10.1038/s41590-026-02544-0

Resource: (BioLegend Cat# 301806, RRID:AB_314188)

Curator: @evieth

SciCrunch record: RRID:AB_314188

What is this?

DOI: 10.1038/s41590-026-02544-0

Resource: (BioLegend Cat# 329950, RRID:AB_2566362)

Curator: @evieth

SciCrunch record: RRID:AB_2566362

What is this?

DOI: 10.1038/s41590-026-02544-0

Resource: (BioLegend Cat# 317426, RRID:AB_571943)

Curator: @evieth

SciCrunch record: RRID:AB_571943

What is this?

DOI: 10.1038/s41590-026-02544-0

Resource: (BioLegend Cat# 302238, RRID:AB_2562097)

Curator: @evieth

SciCrunch record: RRID:AB_2562097

What is this?

DOI: 10.1038/s41590-026-02544-0

Resource: (BioLegend Cat# 302026, RRID:AB_2278418)

Curator: @evieth

SciCrunch record: RRID:AB_2278418

What is this?

DOI: 10.1038/s41590-026-02544-0

Resource: (BioLegend Cat# 353418, RRID:AB_10916518)

Curator: @evieth

SciCrunch record: RRID:AB_10916518

What is this?

DOI: 10.1038/s41590-026-02544-0

Resource: (BioLegend Cat# 362550, RRID:AB_2566059)

Curator: @evieth

SciCrunch record: RRID:AB_2566059

What is this?

DOI: 10.1038/s41590-026-02544-0

Resource: (BioLegend Cat# 321120, RRID:AB_2144930)

Curator: @evieth

SciCrunch record: RRID:AB_2144930

What is this?

DOI: 10.1038/s41590-026-02544-0

Resource: (BioLegend Cat# 304108, RRID:AB_314412)

Curator: @evieth

SciCrunch record: RRID:AB_314412

What is this?

DOI: 10.1038/s41590-026-02544-0

Resource: RRID:AB_2872332

Curator: @evieth

SciCrunch record: RRID:AB_2872332

What is this?

DOI: 10.1038/s41590-026-02544-0

Resource: (BioLegend Cat# 317332, RRID:AB_2561943)

Curator: @evieth

SciCrunch record: RRID:AB_2561943

What is this?

DOI: 10.1016/j.immuni.2026.07.001

Resource: RRID:AB_2927982

Curator: @nmaralla

SciCrunch record: RRID:AB_2927982

What is this?

DOI: 10.1016/j.immuni.2026.07.001

Resource: RRID:AB_2941396

Curator: @nmaralla

SciCrunch record: RRID:AB_2941396

What is this?

DOI: 10.1016/j.immuni.2026.07.001

Resource: (BioLegend Cat# 155871, RRID:AB_2819910)

Curator: @nmaralla

SciCrunch record: RRID:AB_2819910

What is this?

DOI: 10.1016/j.immuni.2026.07.001

Resource: (BioLegend Cat# 155863, RRID:AB_2800694)

Curator: @nmaralla

SciCrunch record: RRID:AB_2800694

What is this?

DOI: 10.1016/j.immuni.2026.07.001

Resource: (BioLegend Cat# 155867, RRID:AB_2800696)

Curator: @nmaralla

SciCrunch record: RRID:AB_2800696

What is this?

DOI: 10.1016/j.immuni.2026.07.001

Resource: RRID:AB_2924484

Curator: @nmaralla

SciCrunch record: RRID:AB_2924484

What is this?

DOI: 10.1016/j.immuni.2026.06.025

Resource: RRID:IMSR_JAX:000664

Curator: @nmaralla

SciCrunch record: RRID:IMSR_JAX:000664

What is this?

DOI: 10.1016/j.immuni.2026.06.025

Resource: RRID:IMSR_JAX:000058

Curator: @nmaralla

SciCrunch record: RRID:IMSR_JAX:000058

What is this?

DOI: 10.1016/j.immuni.2026.06.025

Resource: RRID:IMSR_JAX:007079

Curator: @nmaralla

SciCrunch record: RRID:IMSR_JAX:007079

What is this?

DOI: 10.3390/nu18142307

Resource: (Leica Biosystems Cat# NCL-MEROSIN, RRID:AB_442108)

Curator: @scibot

SciCrunch record: RRID:AB_442108

What is this?

DOI: 10.3390/nu18142307

Resource: (DSHB Cat# N2.261, RRID:AB_531790)

Curator: @scibot

SciCrunch record: RRID:AB_531790

What is this?

DOI: 10.3390/nu18142307

Resource: (DSHB Cat# A4.840, RRID:AB_528384)

Curator: @scibot

SciCrunch record: RRID:AB_528384

What is this?

DOI: 10.3390/nu18142307

Resource: (DSHB Cat# A4.74, RRID:AB_528383)

Curator: @scibot

SciCrunch record: RRID:AB_528383

What is this?

DOI: 10.3390/nu18142307

Resource: (Agilent Cat# M0638, RRID:AB_2249754)

Curator: @scibot

SciCrunch record: RRID:AB_2249754

What is this?

DOI: 10.3390/molecules31142453

Resource: (Agilent Cat# Z0334, RRID:AB_10013382)

Curator: @scibot

SciCrunch record: RRID:AB_10013382

What is this?

DOI: 10.3390/molecules31142453

Resource: (ABclonal Cat# A2572, RRID:AB_2764458)

Curator: @scibot

SciCrunch record: RRID:AB_2764458

What is this?

DOI: 10.3390/molecules31142453

Resource: (Santa Cruz Biotechnology Cat# sc-56053, RRID:AB_781826)

Curator: @scibot

SciCrunch record: RRID:AB_781826

What is this?

DOI: 10.3390/life16071053

Resource: (Thermo Fisher Scientific Cat# 14-9892-82, RRID:AB_10598206)

Curator: @scibot

SciCrunch record: RRID:AB_10598206

What is this?

DOI: 10.3390/life16071053

Resource: RRID:AB_60311

Curator: @scibot

SciCrunch record: RRID:AB_60311

What is this?

DOI: 10.3389/fpsyg.2026.1814576

Resource: R Project for Statistical Computing (RRID:SCR_001905)

Curator: @scibot

SciCrunch record: RRID:SCR_001905

What is this?

DOI: 10.3389/fpsyg.2026.1814576

Resource: RStudio (RRID:SCR_000432)

Curator: @scibot

SciCrunch record: RRID:SCR_000432

What is this?

DOI: 10.3389/fphar.2026.1865172

Resource: GraphPad (RRID:SCR_000306)

Curator: @scibot

SciCrunch record: RRID:SCR_000306

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: Integrated DNA Technologies (RRID:SCR_025813)

Curator: @scibot

SciCrunch record: RRID:SCR_025813

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: Applied Biosystems 7500 Real-Time PCR System (RRID:SCR_018051)

Curator: @scibot

SciCrunch record: RRID:SCR_018051

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (DSMZ Cat# ACC-305, RRID:CVCL_0045)

Curator: @scibot

SciCrunch record: RRID:CVCL_0045

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: RRID:AB_3645670

Curator: @scibot

SciCrunch record: RRID:AB_3645670

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: FlowJo (RRID:SCR_008520)

Curator: @scibot

SciCrunch record: RRID:SCR_008520

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (R and D Systems Cat# MAB139, RRID:AB_2133263)

Curator: @scibot

SciCrunch record: RRID:AB_2133263

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: QIAGEN (RRID:SCR_008539)

Curator: @scibot

SciCrunch record: RRID:SCR_008539

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (Thermo Fisher Scientific Cat# 46-5958-82, RRID:AB_2016714)

Curator: @scibot

SciCrunch record: RRID:AB_2016714

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (Thermo Fisher Scientific Cat# 48-0032-82, RRID:AB_1272193)

Curator: @scibot

SciCrunch record: RRID:AB_1272193

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (BioLegend Cat# 135031, RRID:AB_2564216)

Curator: @scibot

SciCrunch record: RRID:AB_2564216

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (BD Biosciences Cat# 559925, RRID:AB_2869266)

Curator: @scibot

SciCrunch record: RRID:AB_2869266

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (BD Biosciences Cat# 558403, RRID:AB_647201)

Curator: @scibot

SciCrunch record: RRID:AB_647201

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (BD Biosciences Cat# 552878, RRID:AB_394507)

Curator: @scibot

SciCrunch record: RRID:AB_394507

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (BD Biosciences Cat# 557984, RRID:AB_396972)

Curator: @scibot

SciCrunch record: RRID:AB_396972

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (BD Biosciences Cat# 553858, RRID:AB_395094)

Curator: @scibot

SciCrunch record: RRID:AB_395094

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: RRID:AB_394901

Curator: @scibot

SciCrunch record: RRID:AB_394901

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (BD Biosciences Cat# 557235, RRID:AB_396609)

Curator: @scibot

SciCrunch record: RRID:AB_396609

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (BD Biosciences Cat# 553142, RRID:AB_394657)

Curator: @scibot

SciCrunch record: RRID:AB_394657

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (BD Biosciences Cat# 561126, RRID:AB_10611874)

Curator: @scibot

SciCrunch record: RRID:AB_10611874

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (BioLegend Cat# 108913, RRID:AB_492879)

Curator: @scibot

SciCrunch record: RRID:AB_492879

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (BD Biosciences Cat# 553061, RRID:AB_394594)

Curator: @scibot

SciCrunch record: RRID:AB_394594

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (BD Biosciences Cat# 552772, RRID:AB_394458)

Curator: @scibot

SciCrunch record: RRID:AB_394458

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (BD Biosciences Cat# 550992, RRID:AB_398483)

Curator: @scibot

SciCrunch record: RRID:AB_398483

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (BD Biosciences Cat# 562958, RRID:AB_2665409)

Curator: @scibot

SciCrunch record: RRID:AB_2665409

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (Cell Signaling Technology Cat# 2876, RRID:AB_2064177)

Curator: @scibot

SciCrunch record: RRID:AB_2064177

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (Cell Signaling Technology Cat# 5605, RRID:AB_1903938)

Curator: @scibot

SciCrunch record: RRID:AB_1903938

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: Bio-Rad Chemidoc Gel Imaging System (RRID:SCR_019684)

Curator: @scibot

SciCrunch record: RRID:SCR_019684

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: Image Lab Software (RRID:SCR_014210)

Curator: @scibot

SciCrunch record: RRID:SCR_014210

What is this?

DOI: 10.3389/fonc.2026.1874366

Resource: (Sigma-Aldrich Cat# A5441, RRID:AB_476744)

Curator: @scibot

SciCrunch record: RRID:AB_476744

What is this?

DOI: 10.3389/fnins.2026.1838204

Resource: nano@stanford (RRID:SCR_026695)

Curator: @scibot

SciCrunch record: RRID:SCR_026695

What is this?

DOI: 10.3389/fnagi.2026.1824891

Resource: (Abcam Cat# ab53444, RRID:AB_869007)

Curator: @scibot

SciCrunch record: RRID:AB_869007

What is this?

DOI: 10.3389/fnagi.2026.1824891

Resource: (Thermo Fisher Scientific Cat# A-11006, RRID:AB_2534074)

Curator: @scibot

SciCrunch record: RRID:AB_2534074

What is this?

DOI: 10.3389/fnagi.2026.1824891

Resource: (Wako Cat# 019-19741, RRID:AB_839504)

Curator: @scibot

SciCrunch record: RRID:AB_839504

What is this?

DOI: 10.3389/fnagi.2026.1824891

Resource: (Thermo Fisher Scientific Cat# A10042, RRID:AB_2534017)

Curator: @scibot

SciCrunch record: RRID:AB_2534017

What is this?

DOI: 10.3389/fnagi.2026.1824891

Resource: (MMRRC Cat# 034848-JAX,RRID:MMRRC_034848-JAX)

Curator: @scibot

SciCrunch record: RRID:MMRRC_034848-JAX

What is this?

DOI: 10.3389/fnagi.2026.1824891

Resource: RRID:IMSR_JAX:000664

Curator: @scibot

SciCrunch record: RRID:IMSR_JAX:000664

What is this?

DOI: 10.3389/fnagi.2026.1804332

Resource: (MMRRC Cat# 034848-JAX,RRID:MMRRC_034848-JAX)

Curator: @scibot

SciCrunch record: RRID:MMRRC_034848-JAX

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: RRID:AB_3583383

Curator: @scibot

SciCrunch record: RRID:AB_3583383

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: (BD Biosciences Cat# 563410, RRID:AB_2738189)

Curator: @scibot

SciCrunch record: RRID:AB_2738189

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: FlowJo (RRID:SCR_008520)

Curator: @scibot

SciCrunch record: RRID:SCR_008520

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: GraphPad Prism (RRID:SCR_002798)

Curator: @scibot

SciCrunch record: RRID:SCR_002798

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: (BD Biosciences Cat# 560181, RRID:AB_1645235)

Curator: @scibot

SciCrunch record: RRID:AB_1645235

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: (BD Biosciences Cat# 553033, RRID:AB_394571)

Curator: @scibot

SciCrunch record: RRID:AB_394571

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: (Thermo Fisher Scientific Cat# 48-9985-82, RRID:AB_2574139)

Curator: @scibot

SciCrunch record: RRID:AB_2574139

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: (BD Biosciences Cat# 563005, RRID:AB_2737946)

Curator: @scibot

SciCrunch record: RRID:AB_2737946

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: (BD Biosciences Cat# 553104, RRID:AB_394628)

Curator: @scibot

SciCrunch record: RRID:AB_394628

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: (BD Biosciences Cat# 565478, RRID:AB_2739257)

Curator: @scibot

SciCrunch record: RRID:AB_2739257

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: (BD Biosciences Cat# 553312, RRID:AB_398535)

Curator: @scibot

SciCrunch record: RRID:AB_398535

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: (BD Biosciences Cat# 565411, RRID:AB_2734779)

Curator: @scibot

SciCrunch record: RRID:AB_2734779

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: (Proteintech Cat# 21898-1-AP, RRID:AB_2811115)

Curator: @scibot

SciCrunch record: RRID:AB_2811115

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: (R and D Systems Cat# AF2535, RRID:AB_2063012)

Curator: @scibot

SciCrunch record: RRID:AB_2063012

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: (Thermo Fisher Scientific Cat# 14-9760-82, RRID:AB_2572996)

Curator: @scibot

SciCrunch record: RRID:AB_2572996

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: (Thermo Fisher Scientific Cat# MF48000, RRID:AB_10376289)

Curator: @scibot

SciCrunch record: RRID:AB_10376289

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: (Proteintech Cat# 16001-1-AP, RRID:AB_2289842)

Curator: @scibot

SciCrunch record: RRID:AB_2289842

What is this?

DOI: 10.3389/fimmu.2026.1792483

Resource: RRID:IMSR_JAX:027081

Curator: @scibot

SciCrunch record: RRID:IMSR_JAX:027081

What is this?

DOI: 10.3389/fimmu.2026.1758569

Resource: University of California at San Diego Electron Microscopy Core Facility (RRID:SCR_022039)

Curator: @scibot

SciCrunch record: RRID:SCR_022039

What is this?

DOI: 10.14814/phy2.71037

Resource: GraphPad Prism (RRID:SCR_002798)

Curator: @scibot

SciCrunch record: RRID:SCR_002798

What is this?

DOI: 10.14814/phy2.71037

Resource: (RGD Cat# 13508588,RRID:RGD_13508588)

Curator: @scibot

SciCrunch record: RRID:RGD_13508588

What is this?