Fig. 5: Cold-induced PA28γ/PSME-3 prevents aggregation of ALS-related mutant proteins in C. elegans neurons.
a, Western blot with anti-FUS antibody of day 6 adult worms expressing ALS-related FUSP525L mutant variant in neurons. Graph represents the relative percentage of FUSP525L protein levels (corrected for α-tubulin loading control) to 25 °C (mean ± s.e.m., n = 3 independent experiments). b, Cold temperature decreases FUSP525L aggregation in day 6 adult worms (detected by filter trap with anti-FUS antibody). Representative of three independent experiments. c, Western blot of FUSP525L levels upon psme-3 RNAi at different temperatures. Graph represents the relative percentage of FUSP525L levels (corrected for α-tubulin) to 20 °C vector RNAi (mean ± s.e.m., n = 4 independent experiments). d, Filter trap of FUSP525L aggregation upon psme-3 RNAi in day 6 adult worms. Representative of five independent experiments. e, Thrashing movements over a 30-s period at day 3 of adulthood (n = 50 worms from three independent experiments). The box plot represents the 25th–75th percentiles, the line depicts the median and the whiskers show the min–max values. f, Western blot with anti-TDP-43 antibody of day 6 adult worms expressing ALS-related TDP-43M331V mutant variant in neurons. Graph represents the relative percentage of TDP-43M331V protein levels (corrected for α-tubulin) to 25 °C (mean ± s.e.m., n = 3 independent experiments). g, Cold temperature decreases TDP-43M331V aggregation in day 6 adult worms (detected by filter trap with anti-TDP-43 antibody). Representative of three independent experiments. h, Western blot of TDP-43M331V levels upon knockdown of PA28γ/psme-3 at different temperatures. Graph represents the relative percentage of TDP-43M331V levels (corrected for α-tubulin) to 20 °C Vector RNAi (mean ± s.e.m., n = 3 independent experiments). i, Filter trap analysis of TDP-43M331V aggregation upon psme-3 RNAi in day 6 adult worms. Representative of three independent experiments. In all the experiments, worms were shifted to the indicated temperatures after development and RNAi was initiated after development. Statistical comparisons were made by two-tailed Student’s t-test for paired (a,c,f,h) or unpaired samples (e). P value: *P < 0.05, **P < 0.01, ****P < 0.0001; NS, P > 0.05). Significant changes were also significant after correction for multiple testing by FDR (q value < 0.05). Source Data contains for exact P and q values.
