Fig. 3
OPA1 promotes stabilization of ATP synthase oligomers. a Equal amounts (40 µg) of digitonin-solubilized mitochondrial extracts from MAFs of the indicated genotype were separated by BNGE and immunoblotted with anti-ATP synthase subunit α (ATPase) and Succinate Dehydrogenase (SDHA) antibodies. ATPase oligomers (Vo), dimers (Vd), monomers (Vm) and F1 are indicated. b Densitometric analysis of experiments performed as in a. Data represent mean ± SEM of Vo + Vd (oligomeric)/total ATP synthase (Vo + Vd + Vm + F1) from five different experiments. *p < 0.05 in a two-way ANOVA vs. control. c Equal amounts (40 µg) of digitonin-solubilized mitochondrial extracts from MAFs of the indicated genotype were separated by BNGE and immunoblotted with the indicated antibodies (left) or processed for ATPase activity (right). ATPase oligomers (Vo), dimers (Vd), monomers (Vm) and F1 are indicated. d, e Quantitative densitometric analysis of total ATP synthase/CII (SDHA) (d) and of oligomeric/total ATP synthase conformations (e) in experiments as in c. Data show mean ± SEM of at least three independent experiments. *p < 0.05 in a two-way ANOVA versus EV. f Mitochondria from MAFs of the indicated genotypes were treated with recombinant BID as indicated for 30 min, lysed and equal amounts (40 µg) of digitonin-solubilized extracts were separated by BNGE and immunoblotted with the indicated antibodies. g, h Quantitative densitometric analysis of total/CII (SDHA) (g) and of oligomeric/total ATP synthase conformations (h) in experiments as in f. Data are normalized to untreated cells and represent mean ± SEM of at least three independent experiments. *p < 0.05 in an unpaired two-sample Student’s t test versus untreated
