Extended Data Fig. 7: Knockdown or pharmacological inhibition of TRPA1 triggers mutant FUS aggregation at cold temperature.
a, Western blot analysis with anti-FUS antibody of control non-targeting (NT) and TRPA1 shRNA-HEK293 cells expressing mutant FUS(P525L). Graph represents the relative percentage values of FUS protein levels (corrected for β-actin loading control) to 37 °C + NT shRNA (mean ± s.e.m., n = 3 independent experiments). Statistical comparisons were made by two-tailed Student’s t-test for paired samples. P value: *P < 0.05, NS = not significant (P > 0.05). All the significant changes were also significant after correction for multiple testing by FDR approach (q value < 0.05). See Source Data for exact P and q values. b, Filter trap analysis with anti-FUS antibody of NT and TRPA1 shRNA-HEK293 cells expressing mutant FUS(P525L). Knockdown of TRPA1 attenuates the ameliorative effects of cold temperature on FUS(P525L) aggregation levels. Representative of three independent experiments. c, Filter trap analysis with anti-FUS antibody of NT and PSME3 shRNA-HEK293 cells expressing mutant FUS(P525L) treated with 25 µM HC-030031 (TRPA1 antagonist) or DMSO vehicle control for 24 h. Representative of three independent experiments.
