cul-1 RNAi shortens the extended lifespan of daf-2 mutants. (A) Schematic representation of the CUL-1 E3 ligase complex. (B–G) Lifespan curves of long-lived mutants and wild-type worms grown as adults on control empty vector (green lines), daf-16 RNAi (blue lines), and cul-1 RNAi (red lines). (B) daf-2(mu150). Control vector: mean = 33.4 ± 0.7, n = 89/90 (number of animals that died/total; see Materials and Methods); daf-16 RNAi: mean = 17.3 ± 0.5, n = 80/83, P < 0.0001 vs. control; cul-1 RNAi: mean = 25.2 ± 0.4, n = 80/85, P < 0.0001 vs. control. (C) daf-2(e1368). Control vector: mean = 32.7 ± 0.7, n = 58/105; daf-16 RNAi: mean = 22.1 ± 0.3, n = 76/105, P < 0.0001 vs. control; cul-1 RNAi: mean = 25.1 ± 0.4, n = 69/104, P < 0.0001 vs. control. (D) daf-2(e1370). Control vector: mean = 40.8 ± 1.0, n = 72/89; daf-16 RNAi: mean = 22.7 ± 0.5, n = 67/90, P < 0.0001 vs. control; cul-1 RNAi: mean = 31.1 ± 0.9, n = 77/83, P < 0.0001 vs. control. (E) N2. Control vector: mean = 20.7 ± 0.7, n = 67/88; daf-16 RNAi: mean = 16.6 ± 0.5, n = 67/87, P < 0.0001 vs. control; cul-1 RNAi: mean = 18.8 ± 0.6, n = 84/90, P = 0.3 vs. control. (F) glp-1(e2141ts). Control vector: mean = 23.2 ± 0.4, n = 86/87; daf-16 RNAi: mean = 17.8 ± 0.4, n = 79/84, P < 0.0001 vs. control; cul-1 RNAi: mean = 23.9 ± 0.3, n = 74/76, P = 0.4 vs. control. (G) eat-2(ad1116). Control vector: mean = 27.3 ± 0.6, n = 70/96; daf-16 RNAi: mean = 24.8 ± 0.6, n = 68/96, P = 0.005 vs. control; cul-1 RNAi: mean = 25.2 ± 0.5, n = 77/102, P = 0.2 vs. control.