Fig. 6: Mutations at the p53/BCL-2 interface decrease p53-mediated apoptosis.

a Coimmunoprecipitation determines the interaction between full-length p53 and full-length BCL-2. Full-length wild-type or mutant p53 plasmid (Flag-tagged) was co-transfected with full-length BCL-2 plasmid (HA-tagged) into HEK 293 T cells. Cell lysates were immunoprecipitated using anti-Flag beads and subjected to western blotting as indicated. b Caspase 3 and PARP activation detected by western blotting. p53^−/− HCT116 cells stably expressing Ti-p53 or Ti-p53^RRR were seeded in 6-well plates. After 48 h, cell lysates were immunoblotted with antibodies as indicated. c, d Flow cytometry detecting the apoptosis rate. p53^−/− HCT116 cells stably expressing Ti-p53 or Ti-p53^RRR were seeded in 6-well plates. After 48 h, the cells were stained with Annexin V-PE/7-AAD and subjected to flow cytometry. One representative experiment is shown in (c), as three independent replicates were similar. Data are represented as the mean ± SEM of n = 3 independent experiments (d). P values were determined by one-way ANOVA followed by Dunnett’s multiple comparisons test. Source data are provided as a Source Data file.