Fig. 3

ATR inhibitors are more effective in killing breast cancer cells with completed RNF126 rather than corresponding cells with RNF126 knockdown. A Western blot analysis showed that RNF126 knockdown promoted the expression of p-ATR (S428). GAPDH was used as a loading control. B Band intensities were quantified and are presented as bar graphs (Two-way ANOVA, MCF7, upper panel; MDA-MB-231, down panel). C MTT assay showed the effect of the ATR inhibitor (AZD6788) on MCF7 (upper panel) and MDA-MB-231 (down panel) cell viability. Cell viability was expressed as the percentage of viable cells in treated wells relative to the percentage of viable cells in control wells (100% viability). Cells were treated with various concentrations of AZD6788 for 72 h (n = 3, Two-way ANOVA). D Tumor bearing mice were treated with PBS, AZD6738 (50 mg/kg, daily) for 21 days, respectively, Body weight curves of the mice (n = 5 each group). E Tumor growth curves of the mice during the treatment period (n = 5 each group, Two-way ANOVA). F Schematic diagram of tumor on day 42. G Histological observation of metastatic organs includes of lung, liver visualized using H&E staining. H Number of metastases of liver and lung in each group (Two-way ANOVA, MCF7, left panel; MDA-MB-231, right panel). I, J Western blot showed cleaved PARP expression in MCF7 (I) and MDA-MB-231 (J) cells with or without RNF126 depletion. Cells with or without RNF126 shRNA#1 infection were treated with AZD6738 (1 μM) at indicated time points. Band intensities of cleaved PARP were quantified and are presented as bar graphs (Two-way ANOVA). Data are presented as mean ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001. All presented results are from three independent experiments