Fig 1. Homogenates prepared from familial ALS spinal cord tissue induce SOD1 aggregation.
A) SOD1 inclusions in 4 SOD1-FALS cases (A4V, D90A, G93S, I113T) were confirmed by immunohistochemistry. B) Homogenates prepared from human spinal cord tissue were incubated with HEK293FT cells pre-transfected with the indicated reporter protein (G37R, G85R or G93A-based). Cells were imaged 48 h post treatment and analyzed for the presence of inclusions using our algorithm. Representative immunocytochemistry micrographs demonstrate induced aggregation of SOD1G85R-GFP in cells incubated with the indicated homogenate. Arrowheads point towards visible reporter protein inclusions. C) Summary of the effect of FALS, SALS and non-ALS control tissue homogenates on the reporter proteins. Bar graphs represent the percentage of reporter protein in inclusion form out of total reporter protein (area). Statistical significance was established using one way ANOVA followed by Dunnett’s test for multiple comparisons. D) Induced aggregation of the reporter protein using the individual homogenates grouped in (C). Each homogenate was tested 8–16 times with 2 technical repeat per run. *** p < 0.001, * p < 0.05. Scale bar: 40 μm.