To establish that the PI3K-Akt-mTORC1 axis is also constitutively active in patients with the m.3243 A > G mtDNA mutation, we measured p-Akt/Akt and p-S6/S6 by immunofluorescence in a muscle biopsy from a patient with an 82% mutant load of the m.3243 A > G mutation causing MELAS, (not one of the donors used to obtain the fibroblasts; matched H&E and COX staining are shown in Supplementary Fig. 5d). This evidence concerns the gene RPS6 and MELAS.