Fig. 5: RPA1 depletion and cisplatin or Olaparib sensitivity in breast cancer cells.

A RPA1 siRNA knock down in MCF7 cells. Lysates were collected at day3 and day5. B Clonogenic survival assay for cisplatin sensitivity in MCF7 cells control and MCF7_RPA1_KD cells. C Representative photo micrographic images for immunofluorescence staining of 53BP1 in MCF7 control MCF7_RPA1_KD cells treated with Cisplatin (5 μM) for 24 h. D Quantification of 53BP1 nuclear fluorescence by ImageJ software. E Quantification of γH2AX positive cells by flow cytometry. F Cell cycle analysis by flow cytometry. G Annexin V analysis for apoptotic cells in MCF7 control and RPA1_knock down cells treated with 5 μM cisplatin for 24 h. H Clonogenic survival assay for Olaparib sensitivity in MCF7 cells control and MCF7_RPA1_KD cells. I Representative photo micrographic images for immunofluorescence staining of 53BP1 in MCF7 control MCF7_RPA1_KD cells treated with Olaparib (6 μM) for 24 h. J Quantification of 53BP1 nuclear fluorescence by ImageJ software. K Quantification of γH2AX positive cells by flow cytometry. L Cell cycle analysis by flow cytometry. M Annexin V analysis for apoptotic cells in MCF7 control and RPA1_knock down cells treated with 6 μM Olaparib for 24 h. Figures are representative of 3 or more experiments. Statistical analysis was conducted as on GraphPad Prism7 software. To compare between two groups, Student T tests analysis was performed. One-way ANOVA was performed to compare between more than two groups (variances analyses). Two-way ANOVA was used to analyse two variables such as Annexin V analysis and cell cycle analysis. All experiments were expressed as means ± standard deviation S.D. of three independent experiments. Error bars represent standard error of mean between experiments. UN untreated, T treated. *P value <0.05, **P value <0.001, ***P value <0.0001.