Fig. 2: TRPA-1 induces PSME-3 expression via NHR-49 transcription factor in C. elegans at cold temperature.
a, Trypsin-like proteasome activity in day 3-adult wild-type and trpa-1(ok999) mutant worms (mean ± s.e.m. relative slope to 15 °C vector RNAi, n = 4 independent experiments). b, Western blot of PA28γ/PSME-3 in day 6 adult wild-type and trpa-1(ok999) mutant worms. Graph represents the relative percentage values of PSME-3 (corrected for α-tubulin loading control) to 25 °C wild-type (mean ± s.e.m., n = 3 independent experiments). c, qPCR analysis of psme-3 mRNA levels in day 6 adult wild-type and trpa-1(ok999) mutant worms. Graph (relative expression to 25 °C wild-type) represents the mean ± s.e.m. of seven independent experiments. d, psme-3 mRNA levels in day 6 adult control sterile worms upon knockdown of distinct transcriptional regulators involved in cold-induced longevity. Graph (relative expression to 20 °C vector RNAi) represents the mean ± s.e.m. of 9 independent experiments. e, Knockdown of nhr-49 decreases cold-induced trypsin-like proteasome activity in control sterile worms (mean ± s.e.m. relative slope to 20 °C vector RNAi, n = 3 independent experiments). f, pbs-2 mRNA levels in day 6 adult control sterile worms. Graph (relative expression to 20 °C Vector RNAi) represents the mean ± s.e.m. of 9 independent experiments. g, psme-3 mRNA levels in day 6 adult wild-type and trpa-1(ok999) mutant worms. Graph (relative expression to 20 °C Vector RNAi) represents the mean ± s.e.m. of 8 independent experiments. In all the experiments, worms were raised at 20 °C until day 1 of adulthood and then grown at the indicated temperatures until day 3 (a) or 6 (b–g) of adulthood. Statistical comparisons were made by two-tailed Student’s t-test for paired samples. P value: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; NS, P > 0.05. All the significant changes were also significant after correction for multiple testing by FDR approach (q value < 0.05). Source Data contains exact P and q values.
