Fig. 8

Mitochondria morphology and protein synthesis in ALS patient-derived cells. a Mitochondrial morphology in control (Cntl) and C9ALS fibroblasts expressing mito-GFP. BF brightfield. Scale bars = 20 µm. Mitochondrial form factor (a measure of mitochondrial complexity, b) and aspect ratio (an estimate of circularity, c), or both (d) in fibroblasts expressing mito-GFP. Morphological analysis of mitochondrial form factor (e), aspect ratio (f), or both (g) in iPSCs stained with the mitochondrial dye TMRE. *p < 0.05, **p < 0.001 by one-way ANOVA with Tukey’s test. n > 20 cells/group. Results in b−d were combined from four lines each of control and C9orf72 fibroblasts, while e, f were combined from two lines each of control and C9orf72 iPSCs, performed in duplicate. Plots in b, c, e, f show median (horizontal line), interquartile range (box) and maximum/minimum (vertical lines). Graphs in d and g show mean ± standard error. h Bioenergetics analyses demonstrated greater reductions in oxygen consumption rate (OCR) upon addition of 900 nM FCCP, a decoupling agent, to C9ALS and sALS fibroblasts in comparison to controls. n = 8 (Control), 8 (sALS), and 7 (C9ALS) lines/group, as described in Supplementary Table 1. Plot in h shows mean ± 95% confidence interval. i iPSCs from controls and patients carrying C9orf72 mutations displayed elevated protein synthesis by SUnSET. *p < 0.01 by two-sided Kolmogorov−Smirnov test. Inset shows a scatter plot of normalized anti-puromycin counts (mean ± standard error) from control and C9orf72 mutant iPSCs. *p = 0.0129, unpaired t test. n = 2 lines each of control and C9ALS iPSCs, in three separate replicates. j Cumulative distribution function for fractional recovery of mCherry fluorescence in control and C9orf72 iPSCs at 3.5 h. *p < 0.01, two-sided Kolmogorov−Smirnov test. Inset illustrates a scatter plot of fractional recovery (mean ± standard error) in control and C9orf72 mutant iPSCs. *p < 0.01, unpaired t test. n = 2 lines each of control and C9ALS iPSCs, combined from three replicates