Fig. 2: Respiratory defects in patients’ fibroblasts.
From: Mutations of GEMIN5 are associated with coenzyme Q10 deficiency: long-term follow-up after treatment
A–C Oxygen consumption rate (OCR) was measured in patient (P1-3) and control (HDF) fibroblasts. Cells were supplemented with 100 µM of CoQ10 prepared with BSA as vehicle. Non-treated cells were supplemented only with BSA at the same final concentration for 48 h. Arrows indicate the addition of individual inhibitors. Results correspond to the average of six biological replicates ± SD. OL (oligomycin 0.4 μM); FCCP (Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone 0.1 μM); Rot (rotenone 0.1 μM); AntA (antimycin A 0.25 μM). D–F Quantitative data are plotted as the average ± SD (Two-way ANOVA with Sidak’s multiple comparison test). G–I Single (Complex II and III) and combined (Complex II + III) activities of the complexes were measured in fibroblasts homogenates from patients and controls. Results correspond to the average of at least three biological replicates ± SD, normalized by the citrate synthase activity (left axis) or normalized by protein content (right axis) (Unpaired two-tailed t-test). J Citrate synthase activity measured in fibroblast homogenates from patients and controls. Results correspond to the average of at four biological replicates ± SD. K Western blot analysis of VDAC protein levels in whole fibroblast lysates. A stain-free fluorescence signal was used as a loading control. Data correspond to the average of three normalized wells from patients’ fibroblasts expressed as relative expression ± SD. (Unpaired two-tailed t-test). (****p < 0.0001, ***p < 0.001 **p < 0.01 and *p < 0.05).
