Fig. 7: TGFB1 and COL6A1 are neuroprotective in C9ALS/FTD models.
a, qPCR analysis of daw (left panel) and Mp (right panel) transcript levels normalized to Tubulin in WT and GR36 flies. Graph, mean ± s.e.m., n = 8 independent biological replicates, unpaired two-sample Student’s t-test. Genotypes: w; GMR-Gal4/+, w; GMR-Gal4, UAS-GR36/+. NS denotes P > 0.05. b, Stereomicroscopy images of representative 2-day-old adult WT (top panel) or GR36 (bottom panel) Drosophila eyes in absence of or co-expressing daw or Mp RNAi constructs inserted into the second chromosome (II). Genotypes: w; GMR-Gal4/+, w; GMR-Gal4/UAS-daw RNAi, w; GMR-Gal4/UAS-Mp RNAi, w; GMR-Gal4, UAS-GR36/+, w; GMR-Gal4, UAS-GR36/UAS-daw RNAi, w; GMR-Gal4, UAS-GR36/UAS-Mp RNAi. c, Eye size of flies normalized to the mean of the control eye size. Graph, mean ± s.e.m.; n (independent biological replicates, one eye counted per fly) = 20 WT/+, 19 WT/daw RNAi, 23 GR36/+, 11 GR36/daw RNAi; two-way ANOVA, Bonferroni’s multiple comparison. Genotypes: w; GMR-Gal4/+, w; GMR-Gal4/UAS-daw RNAi, w; GMR-Gal4, UAS-GR36/+, w; GMR-Gal4, UAS-GR36/UAS-daw RNAi. d, Eye size of flies normalized to the mean of the control eye size. Graph, mean ± s.e.m.; n (independent biological replicates, one eye counted per fly) = 15 WT/+, 15 WT/Mp RNAi, 12 GR36/+, 10 GR36/Mp RNAi; two-way ANOVA, Bonferroni’s multiple comparison. Genotypes: w; GMR-Gal4/+, w; GMR-Gal4/UAS-Mp RNAi, w; GMR-Gal4, UAS-GR36/+, w; GMR-Gal4, UAS-GR36/UAS-Mp RNAi. e, Percentage cell viability as measured by Alamar Blue following 4-h exposure to 10 µM glutamate. n = 6 control and 6 C9orf72 iPS cell lines. Data points represent average percentage viability from three replicate wells for each condition. Two-way ANOVA with Tukey’s multiple comparison test was used to calculate statistical significance. f, Quantification of the ratio of PI-positive spots to DAPI-positive nuclei (quantification of cell death) following 4-h exposure to 10 µM glutamate. n = 6 control and 6 C9orf72 iPS cell lines. Data points represent average percentage cell death across ten images per well. Two-way ANOVA with Tukey’s multiple comparison test was used to calculate statistical significance.
