Fig. 1: CRC proteins are downregulated in G93A MNs.

a Maximal projections of confocal stacks of VPS35⁺NeuN⁺ putative MNs in the ventral horn of lumbar SCs from WT and G93A mice (30, 60 and 90 days, n = 4 biologically independent mice/time point). Adjacent sections labelled for VPS26 and NeuN are shown in b, (30, 60 and 90 days, n = 4 biologically independent mice/time point). c WBs for VPS35 and β-Actin in lumbar SCs protein extracts from WT and G93A mice at 20, 60, and 100 days (n = 4 biologically independent mice/time point). The histogram shows quantifications (mean ± SD) of normalized VPS35 levels (ratio versus WT), (WT vs. G93A day20: p = 0.007; day 60: p = 0.0015; day 90: p < 0.0001). d Representative WB for VPS26b and β-Tubulin in SCs from WT and G93A mice at day 100. The histogram shows the quantification (mean ± SD) of normalized VPS26b levels (ratio versus WT, n = 5 biologically independent WT mice and n = 6 biologically independent G93A mice, p = 0.002). e Representative WB for VPS29 levels in SCs from WT and G93A mice at day 100. The histogram shows the quantification (mean ± SD) of normalized VPS29 levels (ratio versus WT, n = 6 biologically independent mice/group, p = 0.0019). f quantification of Vps35, Vps26, and Vps29 mRNAs by real-time PCR (ratio versus WT) in lumbar SCs from WT (sampled at day 60, n = 4 biologically independent mice) and G93A mice (30, 60, and 90 days, n = 3 biologically independent mice/time point). Two-way ANOVA followed by Bonferroni multiple comparisons test was used to analyze data of c. Two-tailed Student’s t-test was used to determine the statistical significance in d and e. One-way ANOVA followed by Tukey's Multiple Comparison test was used to analyze data of f. **p < 0.01; ***p < 0.001. Scale bar 80 µm.