FIGURE 5:

Respiratory chain supercomplexes are affected in ssc1-62 mitochondria. (A) Mitochondria from wild-type (WT) and ssc1-62 cells were lysed in digitonin and subjected to centrifugation at 18,000 × g or 125,000 × g, respectively. Supernatant (S) and pellet (P) fractions were separated and analyzed by SDS–PAGE and Western blotting. (B) Mitochondria isolated from WT and ssc1-62 cells grown under permissive conditions were lysed in digitonin and analyzed by blue native electrophoresis followed by Western blotting. III, complex III; IV, complex IV of mitochondrial respiratory chain. (C) WT and ssc1-62 cells were grown under permissive conditions and shifted to 37°C for 32 h. Mitochondria were isolated, and protein complexes were analyzed by blue native electrophoresis followed by Western blotting. TOM, translocase of outer membrane; V, complex V (F₁F_o-ATP synthase). (D) Wild-type and ssc1-62 cells were grown under grown under permissive conditions and shifted to 37°C for 32 h. Isolated mitochondria were lysed under denaturing conditions and analyzed by SDS–PAGE and Western blotting.