Figure 2.

Depletion of mutant p53-R273H induces BMF. Knockdown of mutant p53 induces (a) BMF protein and (b) mRNA expression. MDA-MB-468 cells were transduced with p53 lentiviral shRNA. Protein and mRNA expression were accessed by immunoblot and quantitative RT-PCR, respectively. BMF mRNA expression was normalized against human GAPDH. (c) Ectopic expression of BMF is sufficient to induce apoptosis in MDA-MB-468 cells. Cells were transfected with 5 μg of HA-tagged BMF using X-tremeGENE HP DNA transfection reagent as discussed in the Materials and Methods section. (d–e) Depletion of BMF rescues MDA-MB-468 cells from apoptosis induced by mutant p53 knockdown. Protein lysates and apoptosis were analyzed by (d) immunoblotting and (e) annexin V/7-AAD flow cytometry at 72 h after transduction. Bars represent mean±S.D. of three experiments. * indicates statistical significance (P<0.05) by Student's t-test