Figure 6.

Depletion of NEMF enhances repeat-associated toxicity in a fly model of C9 ALS/FTD and DPR accumulation in human neurons. (A) Representative images of Drosophila eyes expressing (G4C2)28 repeats under the GMR-GAL4 driver in the presence or absence of NEMF at 25°C [BDSC36955 #1 and BDSC25214 #2]. Rough eye phenotypes quantified using an established nominal scoring system are shown as violin plots on the right. Individual flies are represented by single data points. n = 30–33/genotype. (B) (G4C2)28 repeats expressed with a GMR-GAL4 driver at 29°C show decreased eye width that is enhanced by the depletion of NEMF, as quantified on the right. Graphs represent the mean with error bars ± SD, n = 21–24. For A and B, *P< 0.05; ****P≤ 0.0001 by one-way ANOVA with Dunnett's multiple comparison test. (C) Schematic workflow for studies with C9ALS patient-derived iNeurons (iN). (D–F) RNA Expression of NEMF, LTN1, and ANKZF1 transcripts from C9ALS and isogenic control iN lysates. Data represent means with error bars ± SD. n = 3–6/gene, ns = not significant; *P< 0.05 by Student's t-test. (G–I) Quantification of GP by MSD assay from C9ALS and isogenic control iNs treated with lenti-empty vector or lenti-shRNA of NEMF, LTN1 or ANKZF1. Data represent mean ± SD; n = 3–6, ns = not significant; **P≤ 0.01; ***P≤ 0.001 by two-way ANOVA with Sidak's multiple comparison test.