FIGURE 4: Mitochondrial supercomplexes stability is affected in yeast expressing Coq7 phosphomutant versions. Pure mitochondrial samples obtained from the indicated strains were subjected to digitonin solubilization and 200 µg of each sample were analyzed by 1D BN-PAGE.

(A) Coomassie staining of 1D BN-PAGE gels. Molecular weight markers are indicated on the left. On the right side, the three arrows indicate the calculated molecular weight of bands analyzed by densitometry in Figure 4C.

(B) Upper panel, Western blot analysis blotted with antibody against the subunit Cox2 of Complex IV of a 1D BN-PAGE gel obtained in parallel to the Coomassie stained gel shown in Figure 4A using the same original mitochondrial digitonin solubilized samples. Lower panel, analysis by SDS-PAGE and Western blot of mitochondrial samples blotted with antibodies against the subunit Cox2 of Complex IV and porin.

(C) Densitometry analysis of bands indicated with arrows in 4A. The analysis was performed with the software Image Lab 4.0 of Biorad. Band optical density was normalized with the full lane density and compared with the optical density of bands corresponding to the positive control, coq7∆/pNMQ7. Results are representative of a set of two.