Fig. 6.

Mitochondrial superoxide production is due to mutp53-dependent UCP2 inhibition. a Panc1 mutR273H-p53 and AsPC1-p53 null cells were transfected with pRSuper-p53 and vector for mutant p53 ectopic expression, respectively, and their relative controls. In addition, Panc1 cells were co-transfected with siRNA-UCP2, or treated with 150 μM genipin for 24 h; while AsPC1 cells were co-transfected with the UCP2 vector. ROS production, corresponding to DCF fluorescence intensity, was analyzed by a multimode plate reader. *p < 0.05 shCTRL vs shp53 or mock vs R175H or R273H; ^#p < 0.05 shp53 vs shp53 + siUCP2 or shp53 + genipin (Panc1 cells); R175H or R273H vs R175H + UCP2 or R273H + UCP2 (AsPC1 cells). b AsPC1 cells were transfected with the vectors for UCP2 and/or mutant p53 expression. Mitochondrial superoxide production was determined with the MitoSox Red probe by a multimode plate reader. *p < 0.05 CTRL vs R175H or R273H; ^#p < 0.05 R175H or R273H vs R175H + UCP2 or R273H + UCP2. c Panc1 cells were transfected for 48 h with shp53 and/or siUCP2 and their relative negative controls. Cell proliferation was measured by Crystal Violet assay. *p < 0.05 shCTRL vs shp53; shp53 vs shp53 + siUCP2. The experiments are representative of three biological replicates