Fig. 3

Cirp knockout aggravated mitochondrial damage and ROS accumulation via PHD3/HIF-1α axis. A, B Levels of Phd3/β-actin and Hif-1α/β-actin mRNA in kidneys from the sham, DHCA and DHCA + Cirp^−/− groups (n = 5 per group). C Representative western blot images of PHD3 and HIF-1α in renal tissues from the three groups. D The ratio of PHD3 and HIF-1α to β-actin by western blot analysis. E Representative immunofluorescence images of PHD3 and HIF-1α in the renal cortex and medulla from the three groups (original magnification, × 400). F Representative images of extramitochondrial cytochrome c, renal RPA2 and Trx-1 in renal tissues from the three groups by western blot analysis. G The ratio of extramitochondrial cytochrome c, renal RPA2 and Trx-1 to β-actin by western blot analysis. H Representative immunohistochemistry images of cytochrome c in renal cortex and medulla from the three groups (original magnification, × 200). I The cytochrome c-positive cells were quantified by the percentage in 5 randomly selected microscopic vision fields. J Fluorescence intensity of ROS in kidneys from the three groups (per mg protein). ROS, reactive oxygen species. Statistical significance was examined by one-way analysis of variance (ANOVA) followed by the Tukey test. *P < 0.05, **P < 0.01, ***P < 0.001