Fig. 1

Defects in mitochondrial structure in OPA1 R445H iPSC-RGCs. (a) Representative images of the mitochondrial network in R445H and isogenic control neurons stained with Mitotracker dye, deconvoluted with Lightning software. Scale bar, 10 μm. (b-e) MINA analysis of mitochondrial networks. Isogenic, n = 37; R445H, n = 34 cells, sampled from three independent differentiations. (b) and (c), ** = p < 0.01, Mann-Whitney tests, (d) and (e), *** = p > 0.001, Welch’s t-tests. Box plots show median (middle line), 25th-75th percentile (box) and min/max values (whiskers). (f) Representative TEM images of mitochondrial structures in isogenic control and R445H iPSC-RGCs. Scale bar, 2000 nm. (g-i) Quantification of mitochondrial area, roundness, and length, performed in ImageJ. Isogenic, n = 483; R445H, n = 510 ROIs, sampled from three independent differentiations. **** = p < 0.0001, Mann-Whitney tests. Box plots show median (middle line), 25th-75th percentile (box) and min/max values (whiskers). (j & k) Quantification of mitochondrial cristae shape, performed in ImageJ. Isogenic, n = 292; R445H n = 280 ROIs sampled from three independent differentiations. **** = p < 0.0001, Mann-Whitney tests. Box plots show median (middle line), 25th-75th percentile (box) and min/max values (whiskers). (l) Quantification of mitochondria: ER contact sites, performed in ImageJ. Isogenic, n = 350; R445H n = 419 ROIs sampled from three independent differentiations. **** = p < 0.0001, Mann-Whitney tests. Box plots show median (middle line), 25th-75th percentile (box) and min/max values (whiskers). (m) Representative images of Mitotracker-stained mitochondria in TUBB3 + neurites. Scale bar, 5 μm. (n) Quantification of mitochondria length, performed in ImageJ. Isogenic, n = 1,241 ROIs; R445H, n = 625 ROIs, sampled from three independent differentiations. **** = p < 0.0001, Mann-Whitney test. Box plot shows median (middle line), 25th-75th percentile (box) and min/max values (whiskers). (o) Quantification of mitochondria distribution in TUJ1 + neurites. Isogenic/R445H, n = 14 images acquired from three independent differentiations. No significant difference observed between genotypes, p = 0.358, Mann-Whitney test. Scatter plot shows the mean values +/- SD. (p) Quantification of mitochondrial area in TUBB3 + neurites. Isogenic, n = 1,209 ROIs; R445H, n = 590 ROIs, sampled from three independent differentiations. **** = p < 0.0001, Mann-Whitney test. Box plot shows median (middle line), 25th-75th percentile (box) and min/max values (whiskers)