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. 2018 Oct 1;29(20):2386–2396. doi: 10.1091/mbc.E18-04-0227

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© 2018 Möller-Hergt et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

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FIGURE 4: — Mrx15 and Mba1 are required for complex IV accumulation. (A) Steady-state levels of mitochondrial proteins in all indicated strains. Cells were grown in galactose to logarithmic phase, and Western blots probed for proteins necessary for mitochondrial protein biogenesis and complex IV accumulation. (B) Mitochondrial extracts of indicated strains were probed for subunits of complex III (CytB, Qcr7), complex IV (Cox1, Cox13), and complex V (Atp4, F1β). Signal intensities of three independent experiments were quantified and normalized. Significance of observed changes was assessed by a Student’s t test. (C) BN-PAGE of digitonin-solubilized mitochondria from indicated strains. Separated protein complexes were analyzed by Western blotting against subunits of complex III (Rip1), complex IV (Cox1), and complex V (Atp4) (D) Complex III and IV activity measurement. Activity of complex III was followed by measuring cytochrome c reduction, complex IV activity by measuring cytochrome c oxidation at 550 nm. CIII, complex III; CIV, complex IV; CV, complex V; n.s., p > 0.05; **, p ≤ 0.01.