Fig. 1: ALS iPSC-derived MNs exhibit diseased phenotypes.
From: ALS motor neurons exhibit hallmark metabolic defects that are rescued by SIRT3 activation
a Schematic of the MN differentiation protocol. b Immunostaining of wild-type (BJ-iPS, 18a, and GM23720), familial ALS (29d, 47a, and 19f), and sporadic ALS (sALS1, sALS2, and sALS3) iPSC-derived cultures at day 28 indicating the derivation of ISL1+SMI32+ MNs. Cellular nuclei were counterstained with DAPI. Scale bars, 50 μm. c Quantification of ISL1+ MNs from days 25 to 35 demonstrating that wild-type MNs (BJ-iPS, 18a, and GM23720) remain viable while familial and sporadic ALS MNs show significantly reduced survival over time. Quantification of lactate dehydrogenase (LDH) leakage from days 25 to 35 demonstrating that ALS MNs have significantly higher leaked LDH as compared to WT MNs. d qPCR quantification of ER stress transcripts CHOP and spliced XBP1 (sXBP1) in MN cultures at day 28. Fold changes are normalized to expression levels of respective mRNA in BJ-iPS. e Immunostaining of isogenic ALS (BJ-SOD1L144F and BJ-TDP43G298S) iPSC-derived cultures at day 28 indicating the formation of ISL1+SMI32+ MNs. Cellular nuclei were counterstained with DAPI. Scale bars, 50 μm. f Quantification of ISL1+ MNs and leaked LDH derived from BJ-SOD1L144F and BJ-TDP43G298S from days 25 to 35 revealed an accelerated death phenotype and increased LDH leakage similar to that of other ALS lines. g MN cultures derived from BJ-SOD1L144F and BJ-TDP43G298S show upregulation of CHOP and sXBP1 compared to its isogenic control line BJ-iPS. In (d, g), gene expression was normalized to ACTINB and HPRT. ***p < 0.001, ns non-significant; one-way ANOVA, Tukey’s multiple comparisons post hoc test.
