Figure S6.

RCS Are Not Affected by Chronic Loss of Mfn 1 and 2, Related to Figure 6

(A) mtDNA copy number quantification. mtDNA was amplified by RT-PCR from total DNA of the indicated cell lines using specific primers. The mtDNA copy number was normalized to the mtDNA copy number of the correspondent WT cell line. Data are average ± SEM of 5 dependent experiments. ^∗p < 0.05 in a pair sample Student’s t test versus WT.

(B) Representative electron micrographs of cells of the indicated genotype. Bar, 500 μM

(C) Morphometric analysis of cristae width in mitochondria of the indicated genotype. Cristae width was measured in 40 randomly selected mitochondria. Data represent avarage ± SEM of 3 independent experiments.

(D) mtDNA translation assay. The indicated cell lines were metabolic labeled in presence of emetine and lysed after 30 min. Protein samples (40 μg) were separated by SDS-PAGE and the radioactivity was detected in the fixed and dried gels for 3 days. The mtDNA encoded proteins are indicated.

(E) BN PAGE analysis of RCS stability. MEFs of the indicated genotype were metabolically labeled. Equal amounts of protein (100 μg) were separated by BN PAGE and radioactivity was detected in the fixed and dried gels for 3 days. Individual complexes and supercomplexes of the respiratory chain are indicated.

(F) RCS assembly assay. MEFs of the indicated genotype were metabolically labeled and chased for the indicated times. Equal amounts of protein (100 μg) were separated by BN PAGE and radioactivity was detected in the fixed and dried gels for 1 week. Individual complexes and supercomplexes of the respiratory chain are indicated.