Figure 3.

(A) Oxygen Consumption Rate (OCR) is measured before and after the addition of inhibitors. The Seahorse XF Cell Mito Stress Test uses compounds of respiration that target components of the Electron Transport Chain (ETC) in the mitochondria to reveal key parameters of metabolic function. These modulators are ETC inhibitors (oligomycin, FCCP, and a mix of rotenone and antimycin A) which were serially injected to measure ATP (adenosine triphosphate) production, maximal respiration (Max Resp), non-mitochondrial respiration (Non-Mito Resp), proton leak, spare respiratory capacity (SRC) and basal respiration (Basal Resp). (B) The basal energy metabolism of each cell line was assessed by analyzing OCR/ECAR ratios through sequential injections of the inhibitors. (C) Measurement of enzyme activities for the different oxidative phosphorylation system (OXPHOS) complexes in patient and control fibroblasts. FOXRED1 migrated at its predicted size of 54 kDa. CS—Citrate synthase. (D) SDS-PAGE immunodetections of FOXRED1 in control and patient fibroblasts. (M) Marker, (C) Control, and (P) Patient. Blot was cropped from different parts of the same gel to show results from samples of interest. A full-length gel is included in Supplementary Material. (E) One-dimensional Blue-Native polyacrylamide gel electrophoresis (BN-PAGE) analysis of complex I in gel activity and western blot immunodetection showing differences in complex I amount between control (C) and patient (P) fibroblasts. Blot was cropped from different parts of the same gel to show results from samples of interest. A full-length gel is included in Supplementary Material.