Figure 2.

Cytb and Rip1 accumulate in Δcbp3 and Δcbp6-null mutants from the D273-10b lab strain. A, mitochondrial proteins (20 μg) of the indicated mutants were resolved on a 16% SDS-PAGE in the presence of urea 6 m and transferred to PVDF membrane. Western blotting was carried out with anti-Cytb or anti-citrate synthase (as loading control). The asterisk shows an unspecific band. B, mitochondrial proteins (20 μg) of D273-10b and BY4742 mutants were analyzed by SDS-PAGE 12% and Western blotting with the indicated antibodies. C, isolated mitochondria from WT, Δcbp3, and Δcbp6 mutants were first treated with digitonin and sequentially with Triton X-100. After ultracentrifugation, supernatants from both solubilizations were combined. Supernatants (S) and pellet (P) were loaded on a 12% SDS-PAGE and analyzed by Western blotting using antibodies against Cytb, Rip1, and citrate synthase.