Figure 2. OXPHOS complex steady‐state levels and assembly analysis in patient tissues.

• A
  Western blot analysis of OXA1L and OXPHOS complex subunits in patient (P) and control (C1, C2) fibroblasts using VDAC1 and β‐actin as loading controls.
• B
  Western blot analysis of OXA1L and OXPHOS complex subunits in patient and control skeletal muscle. Complex II subunits SDHA and SDHB were used as loading controls.
• C
  Analysis of respiratory complex assembly by Blue Native PAGE in patient fibroblasts solubilised using DDM as a detergent. Antibodies against NDUFB8 (CI), SDHA (CII), UQCRC2 (CIII), COXI (CIV) and ATP5A (CV) were used. * denotes subassembly complex.
• D
  Analysis of respiratory complex assembly by Blue Native PAGE in patient skeletal muscle from the diaphragm (P _dia) and quadriceps (P _quad). Muscle mitochondrial extracts were solubilised using DDM, and antibodies against NDUFB8 (CI), SDHA (CII), UQCRC2 (CIII), COXI (CIV) and ATP5A (CV) were used to detect each complex.
• E
  Western blot analysis of control fibroblasts (C1, C2), patient fibroblasts (P), patient fibroblasts mock transduced with retroviral vector (P+Mock) and patient fibroblasts transduced with retroviral vector containing wild‐type human OXA1L (P+OXA1L). Alpha tubulin was used as a loading control.
• F
  Activities of mitochondrial respiratory complexes I (CI), III (CIII) and IV (CIV), normalised to complex II (CII), in control fibroblasts (white), patient fibroblasts (black) and patient fibroblasts transduced with wild‐type human OXA1L (grey). Mean enzyme activities of control fibroblasts (n = 11) are set to 100%, and error bars represent standard deviation.

Source data are available online for this figure.