Anchorage-independent cell growth was conducted in 6 well plates at least in triplicates. 1% of low temperature melting agarose (Difco) was prepared in sterile MQ water and mixed with 2X DMEM containing 20% FBS kept warm at 37ºC. Both the solutions were mixed in equal proportion to get 0.5% agarose in 1X DMEM and 10% FBS. This solution was kept warm throughout the experiment at 37ºCin a water bath or incubator. To prepare base agar (0.5% agarose), 2 mL of this solution (0.5% agarose+1x DMEM+10% FBS) was dispensed in 6 well plate to spread evenly without any bubbles and allowed to solidify at room temperature at least for 30 min. Cells were harvested and counted using a

Anchorage independent growth

ZFIN_ZDB-GENO-121114-19

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For shame! deny that thou bear'st love to any, Who for thyself art so unprovident. Grant, if thou wilt, thou art beloved of many, But that thou none lovest is most evident; For thou art so possess'd with murderous hate 5 That 'gainst thyself thou stick'st not to conspire. Seeking that beauteous roof to ruinate Which to repair should be thy chief desire. O, change thy thought, that I may change my mind! Shall hate be fairer lodged than gentle love? 10 Be, as thy presence is, gracious and kind, Or to thyself at least kind-hearted prove:     Make thee another self, for love of me,     That beauty still may live in thine or thee

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