Figure 5.

Mutant p53-R273H regulates PI3K/AKT signaling pathway. (a) Knockdown of p53-R273H in MDA-MB-468 cells reduces AKT phorphorylation. MDA-MB-468 cells were transduced with non-targeting (NS) or p53si-1 lentiviral shRNA and lysates were isolated for immunoblotting analysis at 48 and 72 h after transduction. (b) Ectopic expression of mutant p53-R273H, but not p53-R175H, induces AKT phosphorylation. MCF-10A cells were transduced with lentiviral vector or p53si-3 shRNAs targeting the 3′-UTR of the endogenous wild-type p53 followed by brief drug selection (1 μM of puromycin). Pool of p53si-3 stably expressing cells 3 were transfected with vector, p53-R175H or p53-R273H followed by immunoblotting at 72 h after transfection. (c) Ectopic expression of myr-AKT suppresses BMF expression following p53-R273H knockdown in MDA-MB-468 cells. (d) Ectopic expression of myr-AKT abrogated the apoptotic effects induced by p53-R273H depletion in MDA-MB-468 cells. Cells were co-transfected with myr-AKT and p53si-1 for 72 h. Apoptosis was determined using annexin V/7-AAD flow cytometry. Bar represents mean±S.D. of three independent experiments