Extended Data Fig. 1: Cold-induced PSME-3 triggers trypsin-like proteasome activity in young adult C. elegans.
a, Log2-transformed fold changes in label-free quantification (LFQ) of proteasome subunits comparing day 6 adult control sterile fer-15(b26);fem-1(hc17) worms at 15 °C and 20 °C (n = 3). Data from ref. 9, P values calculated by two-tailed Student’s t-test. Worm orthologues of human proteasome subunits were identified with Ortholist2 according to InParanoid8 program34,35. b, Western blot with anti-PSME3 antibody in day 6 control sterile adult worms on psme-3 RNAi. Control sterile worms were raised at 25 °C during development and then grown at the indicated temperatures until day 6 of adulthood. RNAi initiated from day 1 adulthood. α-tubulin loading control. Representative of three independent experiments. c, Trypsin-like activity in day 3-adult control sterile worms (mean ± s.e.m. relative slope to 20 °C Vector RNAi, n = 4 independent experiments). Control sterile worms were raised at 25 °C until day 1 of adulthood and then grown at the indicated temperatures until day 3. d, Trypsin-like activity in day 3-adult wild-type worms (mean ± s.e.m. relative slope to 20 °C Vector RNAi, n = 4 independent experiments). Wild-type worms were raised at 20 °C during development and then grown at the indicated temperatures until day 3 of adulthood. e, Trypsin-like activity in day 1-adult wild-type worms (mean ± s.e.m. relative slope to 20 °C Vector RNAi, n = 4 independent experiments). Worms were raised on RNAi at the indicated temperatures during development. f, Trypsin-like activity in day 6 adult wild-type and trpa-1(ok999) mutant worms at 15 °C (mean ± s.e.m. relative slope to wild-type, n = 7 independent experiments). Worms were raised at 20 °C during development and then grown at 15 °C until day 6 of adulthood. g, Trypsin-like activity in day 6 adult worms at 20 °C (mean ± s.e.m. relative slope to wild-type, n = 7 independent experiments). h, Chymotrypsin-like activity in day 6 adult worms at 15 °C (mean ± s.e.m. relative slope to wild-type, n = 7 independent experiments). In c-h, 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 = not significant (P > 0.05). In c-e, all the significant changes were also significant after correction for multiple testing by FDR (q value< 0.05). Source Data contains exact P and q values.
