Figure 6.

Mutant p53 inhibits Beclin1. (A) AsPC1 cells were seeded in 100‐mm diameter culture dishes and transfected for 48 h with the plasmids coding for mutant p53 proteins (R175H or R273H), or with pCDNA3 as negative control (mock), in the absence or presence of 5 μM everolimus (EVE). Panc1 cells were knocked‐down for mutant p53 expression by 48 h of pRSuper‐p53 vector transfection (shp53). Whole‐cell extracts were used for Western blot analysis using the indicated antibodies. (B and C) Panc1 and SkBr3 cells were seeded in 96‐well plates and transfected with the pRSuper‐p53 vector (shp53) and/or si‐Beclin1 oligos (siBeclin1). Autophagosome formation (B) and cell growth (C) were analyzed after 48 h by the transfection using MDC and crystal violet staining, respectively. These experiments were biological triplicates. The P‐values were calculated with two‐tailed t‐test. Statistical analysis: §p < 0.05 shp53+siBeclin1 vs shp53.