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Fig. 6 | Molecular Medicine

Fig. 6

From: Rapamycin rescues mitochondrial dysfunction in cells carrying the m.8344A > G mutation in the mitochondrial tRNALys

Fig. 6

Mitochondrial respiration and mass assessment in MERRF fibroblasts treated with rapamycin. Wt-Fibroblasts, I-Fibroblasts and H-Fibroblasts were treated four weeks with 20 nM rapamycin. A Representative Western Blot of p-S6, S6 and LC3 proteins. ACTIN was used ad loading control. B Densitometric analyses of downstream target of mTORC1, p-S6 protein. C Densitometric analyses of LC3-II protein. D OCR expressed as pmoles O2/min normalized to cell number, under basal conditions and after injection of oligomycin (O), carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP; F), rotenone (R) and antimycin A (AA). E Basal, ATP-linked and maximal respiration were calculated from OCR traces and reported in the graph. Data are means ± SEM of five experiments for Wt- and H-fibroblasts and of three experiments for I-fibroblasts. F Western Blot analyses of OXPHOS proteins, VDAC and TFAM mitochondrial mass proteins. One representative experiment is shown. Densitometric analyses of OXPHOS G and VDAC and TFAM proteins H. I mtDNA content evaluation by qPCR. J m.8344A > G mutation heteroplasmy evaluation by SNaPshot method. All data are normalized to untreated cells and, if not specifically indicated, are means and SD of six biological replicates for Wt-F, two biological replicates for I-F and four biological replicates for H-F. Statistical analyses were performed using unpaired two-tail T-test. P value: *p < 0.05, **p < 0.01, ***p < 0.001

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