Fig. 2

Tumor necrosis factor receptor-associated protein 1 (TRAP1) regulates the release of apoptosis-associated factors in hypoxic cardiomyocytes. a–d Western blotting and quantitative analysis were used to show the expression of cytochrome c (Cyt c) in the cytosol (a, b) and in the mitochondria (c, d) after Ad_GFP or Ad_TRAP1 transfection with or without hypoxia, indicating the release of Cyt c from mitochondria into the cytosol. *p < 0.05 vs normoxia group; # p < 0.05 vs hypoxia group and hypoxia+Ad_GFP group, respectively. e–h Western blotting and quantitative analysis were used to show the expression of Cyt c in the cytosol (e, f) and in the mitochondria (g, h) after Cont_shRNA or TRAP1_shRNA transfection with or without hypoxia. *p < 0.05 vs normoxia group; #p < 0.05 vs hypoxia group and hypoxia+cont_shRNA group, respectively. i Caspase-3 activity in the myocyte lysates was determined using the caspase-3 Colorimetric Assay Kit after Ad_TRAP1 or TRAP1_shRNA transfection with or without hypoxia. *p < 0.05 vs normoxia group; **p < 0.05 vs hypoxia group and hypoxia+cont_shRNA group, respectively; #p < 0.05 vs hypoxia group and hypoxia+Ad_GFP group, respectively. Data are presented as mean ± SEM. Error bars indicate SEM, p value was analyzed using post hoc Tukey’s tests. The experiment was repeated three times. Ad_TRAP1 adenoviral vector encoding TRAP1, Ad_GFP adenoviral vector encoding green fluorescent protein, TRAP1_shRNA adenoviral shRNA specifically targeting TRAP1, Cont_shRNA adenoviral-mediated expression of scrambled shRNA, GAPDH glyceraldehyde-3-phosphate dehydrogenase