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. 2018 Mar 30;20(5):478–488. doi: 10.1016/j.neo.2018.03.003

Figure 4.

Figure 4

CDK activities are required for the synergistic cytotoxic effect of combination treatment of AZD1775 and AZD6738 in MDA-231 and Hs578t cells.A, MDA-231 and Hs578t cells were treated by DMSO, roscovitine (CDK1/2i), or palbociclib (CDK4/6i) for 24 h, after which DMSO and CDK inhibitors were washed out, and cells were treated with the indicated concentrations of AZD1775 with 100 nM of AZD6738 for another 72 h. Cell viability was detected by CCK8 (n = 3, mean ± SD). B, MDA-231 cells were pretreated with 2 μM of CDK1/2 inhibitor roscovitine, or 2 μM of CDK4/6 inhibitor palbociclib for 6 h before the medium was changed with that containing 100 nM of AZD1775 and 500 nM of AZD6738 for 24 h. Cells after treatment were probed with anti-γH2AX and anti-RAD51 antibodies and the respective images are shown. Scale bar: 10 μm. C, Quantitative data of the pan-nuclear γH2AX after treatment in Figure 4B are presented (n = 2, mean ± SD) (**P < .01 and ***P < .001).