Figure 3.

Western blot analysis of mitochondrial proteins. (a) Twenty microgram of total cellular proteins from various cell lines were electrophoresed through a denaturing polyacrylamide gel, electroblotted and hybridized with four respiratory complex subunits in mutants and controls with VDAC as a loading control. p.MT-ND5, subunits 5 of the reduced nicotinamide–adenine dinucleotide dehydrogenase; p.MT-CO2 indicate subunits 2 of cytochrome c oxidase; p.MT-Cytb, apocytochrome b; and p.MT-ATP6, subunit 6 of the H + -ATPase. (b) Quantification of total mitochondrial protein levels. The levels of mitochondrial proteins in 3 mutant cell lines and 3 control cell lines were determined as described elsewhere²⁷. The values for the mutant cell lines are expressed as percentages of the average values for the control cell lines. The calculations were based on three independent determinations. Graph details and symbols are explained in the legend to Fig. 2. (c) Quantification of 4 respiratory complex subunits. The levels of p.MT-ND5, p.MT-Cytb, p.MT-CO2 and p.MT-ATP6 in 3 mutant cell lines and 3 control cell lines were determined as described elsewhere²⁷.