Research Paper Volume 8, Issue 9 pp 2100—2126

Figure 4. Cell cycle inhibits SKN-1 and DAF-16-driven stress response. (A) Activation of the Pgst-4::GFP reporter in L4/young adult animals that were exposed to cye-1(RNAi), cdk-2(RNAi), or control beginning from L1. (B) Analysis of Pgst-4::GFP expression in skn-1(zu67) mutants. In (A) and (B) induction of the Pgst-4::GFP reporter in the intestine was quantified as low (L), medium (M) and high (H) (see Supplementary Figure 3C). Pooled data from 2 experiments. p-values were calculated by the Chi-square test. ***p < 0.001. ns, not significant. n, number of worms analyzed. (C) Induction of endogenous SKN-1 target gene expression in response to cye-1(RNAi) analyzed by qPCR. RNAi was performed from L4 to day four of adulthood. Data are presented as fold change compared to wild-type on control(RNAi) averaged from at least three independent experiments, error bars represent SEM. p-values were derived from a student’s t-test. *p< 0.05. **p<0.01. (D) cye-1(RNAi) induces expression of Psod-3::GFP (see also Supplementary Figure 4). (E) Analysis of Psod-3::GFP expression in daf-16(mu86) mutants. In (D) and (E) Psod-3::GFP quantification with high (H), medium (M), and low (L) scoring. Pooled data from at least 2 experiments. p-values were calculated by the Chi-square-test. ***p < 0.001. ns, not significant. n, number of worms analyzed. (F) DAF-16 target gene expression assayed by qPCR. Worms were exposed to cye-1(RNAi) or empty vector control during adulthood. The nnt-1, stdh-1 and gpd-2 genes have been shown previously to be upregulated by germline removal. Data are mean ± SEM. p-values were derived from a student’s t-test. *p< 0.05. **p<0.01.