Figure 1 | Scientific Reports

Figure 1

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 1

MCAD-deficient patient fibroblasts have reduced steady-state levels of OXPHOS complexes and supercomplexes. Mitochondria were isolated from control (C1 and C2) or MCAD-deficient patient (P1 and P2) fibroblasts for SDS-PAGE and BN-PAGE Western blot analysis. (A) No mature MCAD protein is detectable in P1 or P2 patient mitochondria by SDS-PAGE (the OXPHOS complex II subunit SDHA is shown as a loading control). BN-PAGE revealed that the steady-state levels of mature OXPHOS complex IV (detected with an anti-COI antibody) (B), complex I (anti-NDUFA9 antibody) (C), the complex III dimer (CIII2) and the CIII2/CIV supercomplex (anti-UQCRC1 antibody) (D) and the CI/CIII2/CIV supercomplex (anti-NDUFA9 antibody) (E) were all reduced in P1 and P2 patient mitochondria compared to C1 and C2 controls respectively. (F) Steady-state levels of OXPHOS complex V (anti-ATP5A antibody) were no different in P1 and P2 patient mitochondria compared to controls. Mitochondria were solubilised in TX-100 (B–D,F) or in digitonin (E). Quantitation is relative to the steady-state levels of OXPHOS complex II (anti-SDHA antibody) (n = 3).

Back to article page