RNAi of cul-1 complex genes affects DAF-16 transcriptional activity. (A) daf-16(mu86); daf-2(e1370); Pdaf-16::daf-16AM::gfp worms were subjected to RNAi treatment from day 1 of adulthood. Empty vector control (green curve): mean = 34.6 ± 0.6, n = 94/102; daf-16 RNAi (blue curve): mean = 22.7 ± 0.7; n = 84/90, P < 0.0001 vs. control; cul-1 RNAi (red curve): mean = 24.3 ± 0.3, n = 95/104, P < 0.0001 vs. control. (B–G) daf-2(e1370); Psod-3::gfp worms subjected to RNAi of cul-1 complex genes from the L4 stage through day 2 of adulthood. sod-3 expression is increased in daf-2 mutants (B) in a daf-16-dependent manner (C). This increased expression is significantly reduced in RNAi of cul-1 (D), skr-1/2 (E), and lin-23 (F). A milder, variable effect was observed on phi-3 RNAi (G). Data from multiple trials are shown in SI Table 5. Bar graph representation is found in SI Fig. 7.