Fig. 3.

Mutant p53 downregulates UCP2 and PGC-1-α. a Panc1 mutR273H-p53 and AsPC1-p53 null cells were transfected with pRSuper-p53 vector and with plasmids for the ectopic expression of mutant p53-R273H or its relative negative control (CTRL). Gene expression analysis of the p53, UCP2, and PGC-1α was performed by RT-qPCR and was normalized to GAPDH mRNA. *p < 0.05. b H1299 p53-null cells stably expressing p53-R273H (clone H1) and respective mock control (clone C9) were used to analyze PGC-1α and UCP2 expression by RT-qPCR, normalized to GAPDH mRNA, and ROS levels with DCF probe. The western blotting was performed using 50 μg of whole-cell extracts and probed with p53 and vinculin antibodies *p < 0.05. c Panc1 mutR273H-p53 and AsPC1-p53 null cells were treated with 40 µM RITA and 20 µM CP-31398 for 48 h and gene expression analysis of UCP2 was performed by RT-qPCR and normalized to GAPDH mRNA. *p < 0.05. d Western blotting analysis of Panc1 mutR273H-p53 and AsPC1-p53 null cells transfected with the indicate plasmids. Western blotting was performed using 50 μg of whole-cell extracts, probed with the indicated antibodies and quantified with ImageJ software. e Panc1 mutR273H-p53 cells were transfected with pRSuper-p53 vector and siRNA-PGC-1α and relative controls and gene expression analysis of UCP2 was performed by RT-qPCR and normalized to GAPDH mRNA. *p < 0.05 shCTRL vs shp53; ^#p < 0.05 shp53 vs shp53 + siPGC-1α. The experiments are representative of three biological replicates