Western blotting analysis of mitochondrial proteins.
A, 20 μg of total cellular proteins from lymphoblastoid cell lines were electrophoresed through a SDS-polyacrylamide gel, electroblotted, and hybridized with seven respiratory complex subunits in mutant and control cells with VADC 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; A6, subunit 6 of the H+-ATPase; and CYTB, apocytochrome b. B, quantification of mitochondrial protein levels. Average content of CO2, ND1, ND4, ND5, ND6, A6 and CYTB per cell, normalized to the average content of VADC per cell in mutant cell lines and controls. The values for the mutant cell lines are expressed as percentages of the average values for the control cell lines. The horizontal dashed lines represent the average value for each group. The calculations were based on three independent determinations. Graph details and symbols are explained in the legend to Fig. 4.