FIG 5.

Western blot analysis of mitochondrial proteins. (A) Twenty micrograms of total cellular proteins from various cell lines was electrophoresed through a denaturing polyacrylamide gel, electroblotted, and hybridized with seven respiratory complex subunits in mutant and control cells with TOM20 as a loading control. CO2, subunit II of cytochrome c oxidase; ND1, ND4, ND5, and ND6, subunits 1, 4, 5, and 6 of the reduced nicotinamide-adenine dinucleotide dehydrogenase; ATP6, subunit 6 of the H⁺-ATPase; CYTB, apocytochrome b. (B) Quantification of mitochondrial protein levels. The levels of mitochondrial proteins in three mutant cell lines and three control cell lines were determined as described elsewhere (28). (C) Quantification of seven respiratory complex subunits. The levels of CO2, ND1, ND4, ND5, ND6, ATP6, and CYTB in three mutant cell lines and three control cell lines were determined as described elsewhere (32). Graph details and symbols are explained in the legend of Fig. 3.