Fig. 3.

Single, double, and triple Opa1 C-terminal Cys mutants generate the 180 KDa Opa1 oligomer upon H₂O₂ treatment.

(A) Equal amounts of proteins (15 μg) from MEFs of the indicated genotypes treated where indicated with 1 mM H₂O₂ for 30 min were separated by non-reducing SDS-PAGE and immunoblotted using the indicated antibodies. Asterisks indicate the ∼180 KDa Opa1 reduction-sensitive oligomer.

(B) Representative confocal images of mitochondrial morphology in WT and Opa1^−/− MEFs cotransfected with mtRFP and the indicated plasmids for 24 h. Scale bar: 20μm.

(C) Quantification of mitochondrial major axis length. Boxes represent 25th-75th percentiles with median values, whiskers 10th-90th percentiles. Values of the individual (4–13, at least 50 cells/experiment) independent experiments performed as in B are plotted as open dots. *. P < 0.05 in a one way ANOVA with Tukey's means comparison between Opa1^WT and the indicated Opa1 mutants.