Figure 5.
Investigation of the role of BAK, BAX and p53 in the activity of ABT-263 and SAR405838 in the RS4;11 cell line. A, Immunoblots illustrating the efficiency of the shRNA-lentiviral approach in RS4;11 and RS4;11/G3M1 cells. B, BAX or BAK was knocked down by shRNA in the RS4;11 parental cell line and cells were treated with ABT-263 for 4 days for cell viability analysis by a WST assay or treated with ABT-263 for 24 h for apoptosis analysis by flow cytometry with Annexin V/P.I. double staining (***, P <0.001; ****, P <0.0001; t test). C, BAK or BAX was efficiently knocked down by shRNA in the RS4;11 parental cell line or BAK was knocked down in G3M1 subline with a very low level of BAX. Cells were treated with SAR405838 for 4 days for cell viability analysis using a WST assay. Apoptosis analysis by flow cytometry with Annexin V/P.I. double staining. RS4;11/shControl, G3M1/shControl and G3M1/shBAK cells were treated with SAR405838 for 24 h (*, P <0.05; **, P <0.01; t test). D, p53 was knocked down by shRNA in the RS4;11 parental cell line and the cells were treated with ABT-263 for 4 days for cell viability analysis in a WST assay or for 24 h for apoptosis analysis by flow cytometry with Annexin V/P.I. double staining.