Fig. 2

Mitochondrial functionality of mutant TARDBP and C9ORF72 fibroblasts in galactose medium. a Measurement of mitochondrial membrane potential (MMP) by flow cytometry analysis of TMRM-positive fibroblasts (4 healthy controls, 3 TARDBP p.A382T (mTDP-43) and 4 C9ORF72 (mC9ORF72). Median ± SEM, n = 3 different DIV; one-way ANOVA with Dunnett’s multiple comparison test, *p < 0.05; **p < 0.01. b Evaluation of ATP content by the ATPlite kit. Mean ± SEM, n = 3 different DIV; one-way ANOVA with Dunnett’s multiple comparison test, **p < 0.01. c. Analysis of oxygen consumption rate in basal condition (OCR-B), upon treatment with oligomycin (OCR-O) and with the oxidative phosphorylation uncoupler FCCP (OCR-F). Data are presented as mean ± SEM; n = 5 different DIV; two-way ANOVA with Bonferroni post-tests; *p < 0.05 and **p < 0.01 vs. basal. d Measurement of the respiratory electron transport system activity (complexes I-IV) and of succinate dehydrogenase (SDH) in 3 CTRL, 3 mTDP-43 and 4 mC9ORF72 fibroblasts. Values are expressed as nmol/min/mg protein and normalised with citrate synthase activity (mean ± SEM, one-way ANOVA with Dunnett’s multiple comparison test; *p < 0.05). e Mitochondrial ROS detection by flow cytometry analysis after incubation with MitoSOX™ Red reagent. Mean ± SEM; n = 3 different DIV; one-way ANOVA with Dunnett’s multiple comparison test; **p < 0.01. f Cellular ROS detection after incubation with 2′,7′-dichlorodihydrofluorescein (DCF). Fluorescence intensity was normalized for cell number. Mean ± SEM; n = 3 different DIV; one-way ANOVA with Dunnett’s multiple comparison test; **p < 0.01