Fig. 6

RPS3/RAD50 knockdown sensitizes K464R mutant cells to Olaparib. (A-B) SKOV3 (A) and A2780 (B) MRE11^K464R cells were transfected with siRAD50 or siRPS3 alone or combine for 24 h and then treated for 96 h with indicated doses of Olaparib and viability was measured by CCK8. The expression of scramble siRNA (siNC) was used as control. (C-D) Cells were transfected with siRAD50 or siRPS3 alone or combine for 24 h and then treated with or without Olaparib for 10 days. The expression of siNC was used as control. Representative pictures of clonogenic assay in A2780 MRE11^K464R cells (C). The mean clones of ROI are presented (D). (E-F) SKOV3 (E) and A2780 (F) MRE11^K464R cells were transfected with siRAD50 or siRPS3 alone or combine for 24 h and then treated for 48 h and then subjected to Comet analysis. DNA damage is quantified as percent DNA in tails. The expression of siNC was used as control. Each group represents at least 150 cells counted. (G-H) SKOV3 (G) and A2780(H) MRE11^K464R cells were transfected with siRAD50 or siRPS3 alone or combine for 24 h and then treated with or without Olaparib for 48 h and then stained for γH2AX (green) and DAPI (blue), and γH2AX foci-positive cells were quantified (below). Each group represents at least 150 cells counted. Scale bar, 20 μm. Data are presented as mean values ± SEM from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns, not significant, as determined by the unpaired two-tailed Student’s t-test