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A
Representative western blots of whole‐cell lysates from MCF7 cells treated with palbociclib (1 μM) for 1, 4 or 7 days, or treated identically, and then washed out for the indicated times to allow cells to enter S‐phase.
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B
Analysis of adjusted relative density from three independent western blot experiments. Bars display mean values ± SD. Significance determined by unpaired Student's t‐test comparing treated target protein to asynchronous target control (*< 0.01, **< 0.001, ***< 0.0001).
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C, D
Quantification of the nuclear morphologies (C) or γH2AX‐positive DNA damage foci (D) from MCF7, MCF7 p53 KO or T47D cells that were treated with palbociclib (1 μM) for 0, 1, 4 or 7 days and then analysed 72 h after drug washout. Either 100 cells (nuclear morphology) or 50 cells (γH2AX) were scored per condition per experiment and bar graphs represent mean data + SEM from three experiments.
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E
Cumulative mitotic entry of cells following washout from 0, 1, 4 or 7 days treated of palbociclib (1 μM). A total of 50 cells were quantified per experiments and graphs display mean ± SEM from three experiments.
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F
Quantification of colony forming assays of MCF7, MCF7 p53 KO and T47D cells treated with CDK4/6 inhibitor for 0, 1, 4 or 7 days and then grown at a low density without palbociclib for 14–21 days. Bar graphs display mean data + SEM from 4 to 5 experiments.
