Induction of p65 enhanced the p53-mediated cell death response to DOX in PANC1 cells. Cells were transiently transfected with or without over-expression of p53 alone or p53 and p65 together followed by DOX treatment (2 μg/ml) for 24 h. (A) The level of p53 or p65 protein was detected by Western blotting. (B) Cell survival was examined by CCK8 kit. Error bar indicates the standard error of the mean of three independent experiments. (C)The nuclear or cytoplasmic p21 protein (above) or pro-caspase-3 (bottom) were detected by Western blotting. GAPDH was used as controls. Whole cell extract was subjected to immunoprecipitation with anti-p21 antibody and then immunoblotted with anti-pro-caspase-3 antibody (middle). (D) Active-caspase-3 and active-caspase-8 protein were detected by Western blotting. GAPDH was used as controls.