Fig. 3

Analysis of autophagy and mitochondria biogenesis in mutant TARDBP and C9ORF72 fibroblasts in galactose medium. a Representative WB analysis of p62 and LC3II/I in lysates of control (CTRL) and mutated TARDBP (mTDP-43) fibroblasts (left panel). Tubulin was used for sample normalization. Densitometric analyses of WB data are shown in the right panels; the ratio between lipidated (II) and unlipidated (I) protein is shown for LC3 (mean ± SEM of 3 controls and 3 mTDP-43 patients; n = 3 different DIV; unpaired t-test, **p < 0.01). b Representative WB analysis of p62 and LC3II/I in lysates of CTRL and mutated C9ORF72 (mC9ORF72) fibroblasts (left panel). Tubulin was used for sample normalization. Densitometric analyses of WB data are shown in right panels (mean ± SEM of 3 controls and 4 mC9ORF72 patients, n = 3 different DIV; unpaired t-test, *p < 0.05). Q-PCR analysis of c SQSTM1/p62 and d MAP1LC3 mRNA content in CTRL, mTDP-43 and mC9ORF72 fibroblasts. Fold change values were calculated versus CTRL fibroblasts (mean ± SEM; n = 3 different DIV; one-way ANOVA with Dunnett’s multiple comparison test). e Quantification of mitochondrial DNA content by Q-PCR of the mitochondria-encoded NADH dehydrogenase 5 (MT-ND5) gene. The genomic ribonuclease P gene was used for data normalization (mean ± SEM; n = 3 for each experimental group; one-way ANOVA with Dunnett’s multiple comparison test). f Representative WB analysis of PGC1α in lysates of controls (CTRL), mTDP-43 (left panel) and mC9ORF72 (right panel) fibroblasts. Tubulin was used for sample normalization. Densitometric analyses of WB data are shown in the lower left and right panels (mean ± SEM; n = 3 different DIV; unpaired t-test, *p < 0.05). g Measurement of mitochondrial mass (MM) by flow cytometry analysis of Mitotracker green (MTG)-positive fibroblasts (4 healthy controls, 3 mTDP-43 and 4 mC9ORF72). Median ± SEM; n = 3 different DIV; one-way ANOVA with Dunnett’s multiple comparison test; **p < 0.01