Figure 4 | Scientific Reports

Figure 4

From: Loss of the Mitochondrial Fatty Acid β-Oxidation Protein Medium-Chain Acyl-Coenzyme A Dehydrogenase Disrupts Oxidative Phosphorylation Protein Complex Stability and Function

Figure 4

Incorporation of newly-translated mtDNA-encoded subunits into the OXPHOS complexes and the OXPHOS supercomplex is disrupted in MCAD knockout cells. MtDNA-encoded-proteins were radiolabeled in the presence of cycloheximide and chased for 0, 3 or 24 h. (A) SDS-PAGE showing similar amounts of newly-translated complex I (ND1, ND2, ND3, ND4L, ND5 and ND6), complex III (cyt b), complex IV (COI, COII and COIII) and complex V (ATP6 and ATP8) subunits in both 143B control (CON) and 143B MCAD knockout (KO) mitochondria. (B) BN-PAGE analysis of mitochondria solubilised in TX-100 shows that the amount of newly-translated mtDNA-encoded proteins incorporated into complex I (CI), the complex III dimer (CIII2) and complex IV (CIV) after 24 h chase is less in MCAD KO mitochondria than CON mitochondria. Levels of complex V were not different (C) BN-PAGE following solubilisation in digitonin. Reduced levels of newly-translated subunits in the CI/CIII2/CIV supercomplex are evident in MCAD KO mitochondria compared to control (CON). (D) Quantitation of OXPHOS complex and supercomplex levels. Data is mean ± s.d., n = 3. *p < 0.05, **p < 0.01.

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