Figure 5. cbs-1 and other factors modulate the long lifespan induced by adult GSCs at low temperatures.
a, Heatmap depicting the log2-fold change of the differentially expressed proteins in both iff-1 RNAi-treated worms at 15°C and empty vector (EV) RNAi-treated worms at 20°C when compared with EV RNAi-treated worms at 15°C. fer-15(b26);fem-1(hc17) control strain was raised at the restrictive temperature (25°C) during development to obtain sterile worms with a proliferating germline, which were then shifted to the indicated temperatures and RNAi treatment until day 6 of adulthood. Statistical comparisons were made by two-tailed Student’s t-test (n= 3, p-value <0.05 was considered significant). Lifespan experiments upon knockdown of the indicated proteins are presented in the following panels. Supplementary Fig. 17 presents the heatmap with the full list of significantly changed proteins. For lifespan data of the other differentially expressed proteins, please see Supplementary Table 1. b, CCT subunits are required for cold-induced longevity (15°C) in wild-type worms (Empty vector RNAi mean ± s.e.m: 28.26 ± 0.69; cct-2 RNAi: 18.58 ± 0.71, P<0.0001; cct-5 RNAi: 17.83 ± 0.68, P<0.0001; cct-6 RNAi: 16.15 ± 0.66, P<0.0001; hint-1 RNAi: 27.15 ± 0.69, P=0.1722). c, The hsp-1 chaperone is necessary for cold-induced longevity (15°C) (Empty vector RNAi mean ± s.e.m: 25.49 ± 0.62; hsp-1 RNAi: 19.84 ± 0.49, P<0.0001; fkb-6 RNAi: 24.10 ± 0.73, P=0.2535). d, The RNA polymerase II ama-1 regulates longevity of wild-type worms at 15°C (Empty vector RNAi mean ± s.e.m: 28.54 ± 0.68; ama-1 RNAi: 23.67 ± 0.42, P<0.0001; ttr-6 RNAi: 27.95 ± 0.65 P=0.4734). e, Loss of cbs-1 decreases longevity of wild-type worms at 15°C (Empty vector RNAi mean ± s.e.m: 28.26 ± 0.69; cbs-1 RNAi: 24.54 ± 0.69, P=0.0005; cal-5 RNAi: 28.09 ± 0.78, P=0.8688; nlp-24 RNAi: 27.16 ± 0.78, P=0.0664). f, Knockdown of cct subunits, hsp-1, ama-1 or cbs-1 decreases the long lifespan phenotype induced by 10°C in wild-type worms (P<0.0001). Empty vector RNAi mean ± s.e.m: 43.31 ± 1.39; cct-2 RNAi: 26.36 ± 0.02; cct-5 RNAi: 26.76 ± 0.95; hsp-1 RNAi: 30.17 ± 0.82; ama-1 RNAi: 35.37 ± 0.87; cbs-1 RNAi: 33.57 ± 0.93. g, Knockdown of cct subunits, hsp-1, ama-1, or cbs-1 does not shorten lifespan at 20°C. Empty vector RNAi mean ± s.e.m: 16.34 ± 0.37; cct-2 RNAi: median= 15.97 ± 0.41, P=0.5386; cct-5 RNAi: 17.01 ± 0.48, P=0.1106; hsp-1 RNAi: 17.10 ± 0.39, P=0.1571; ama-1 RNAi: 17.02 ± 0.35, P=0.4403; cbs-1 RNAi: 16.26 ± 0.43, P=0.9592. h, Knockdown of cct subunits, hsp-1, ama-1, or cbs-1 does not shorten lifespan at 25°C. RNAi was initiated at day 1 of adulthood. Empty vector RNAi mean ± s.e.m: 13.95 ± 0.37; cct-2 RNAi: 13.20 ± 0.31, P=0.0695; cct-5 RNAi: 13.43 ± 0.29, P=0.0472; hsp-1 RNAi: 13.83 ± 0.36, P=0.7389; ama-1 RNAi: 14.07 ± 0.36, P=0.8221; cbs-1 RNAi: 14.53 ± 0.44, P=0.1069. In all the lifespan experiments, RNAi was started at day 1 of adulthood (P-values: two-sided log-rank test, n= 96 worms/condition). See Supplementary Data 3 for statistical analysis and replicate data of lifespan experiments.