Fig. 3.

Cytochrome c oxidase subunit II (COXII) knockdown prevented the antiapoptotic effect of Ad_TRAP1 on hypoxic cardiomyocytes. a Representative images of cardiomyocyte apoptosis were detected by TUNEL staining after transfection with Ad_TRAP1, COXII_shRNA, or both, with or without hypoxia. Scale bar = 50µm. b Apoptosis rates of cardiomyocytes were quantitatively analyzed in each treatment group. c, d Western blotting (c) and quantitative analysis (d) were performed to detect cytochrome c (Cyt c) levels in the cytosol. GAPDH served as an internal control. e Caspase-3 activity in the cardiomyocytes was determined using the caspase-3 Colorimetric Assay Kit in each treatment group. *p < 0.05 vs normoxia group; **p < 0.05 vs. hypoxia group and hypoxia+Ad_GFP group, respectively; #p < 0.05 vs hypoxia+Ad_TRAP1 group and hypoxia+Ad_TRAP1+Cont_shRNA 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, COXII_shRNA adenoviral shRNA specifically targeting COXII, Cont_shRNA adenoviral-mediated expression of scrambled shRNA, GAPDH glyceraldehyde-3-phosphate dehydrogenase, TUNEL terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling