Fig. 5: Effect of AOX on OXPHOS, NAD(H) redox status, and mitochondrial ROS production in Bcs1l^p.S78G;mt-Cyb^p.D254N livers.

a Phosphorylating respiration in liver mitochondria in the presence of malate, glutamate, pyruvate, and ADP (CI-linked respiration). b Phosphorylating respiration of liver mitochondria in the presence of malate, glutamate, pyruvate, succinate, and ADP (CI&CII-linked respiration). c CI&CII-linked respiration after inhibition of AOX with 50 µM propyl gallate. d, e Blue-native PAGE analysis of CIII assembly from digitonin-solubilized liver mitochondria. The RISP subunit is present only in the fully assembled CIII dimers. f ATP production in liver mitochondria during CI-linked respiration. g Membrane potential of liver mitochondria during CI-linked phosphorylating respiration (OXPHOS). The subsequent addition of ATP-synthase inhibitor, oligomycin A, and the uncoupler, SF-6847, were used to obtain maximal and dissipated membrane potential, respectively. h Enzymatic determination of ATP in liver tissue. i Hepatic NADH-to-NAD⁺ ratio. j, k Succinate and fumarate levels in liver. l Measurement of H₂O₂ production in isolated liver mitochondria. The mitochondrial antioxidant systems and carboxylesterases were blocked as described in the Materials and Methods. m, n Non-reducing Western blot of PRDX3 (m) and PRDX1 (n). One sample treated with β-mercaptoethanol (βME) served as a control to verify the specificity of bands corresponding to peroxiredoxin dimers. All data are from 1-month-old mice. Statistics: d, χ²-test on binarized data (detected or not detected); all other data, one-way ANOVA followed by the selected pairwise comparisons (Welch’s t-statistics). The error bars represent 95% CI of mean. All data points derive from independent mice. Source data are provided as a Source Data file.