Figure 4.

MEKK−3 and NSY−1 are indispensable for the induction of SKN−1 target genes. (a) Intestinal expression of GCS-1 labeled with GFP. GCS-1 transgenic young adult worms were fed bacteria that carry RNAi targeting either mekk-3, nsy-1, or both mekk-3 and nsy-1 RNAi for 24 h. The L4440 vector without insertion in bacteria was used as a control. Fluorescent images were captured 6 h after the transfer of worms fed RNAi bacteria on 7.5 mM t-BOOH plates. The white arrows indicate GCS-1::GFP. (b) Intestinal GCS-1::GFP expression was scored in young adult worms (n = 30 in each group). (c) Quantitative PCR analyses of gcs-1 (n = 3). Total RNA extraction from GCS-1 transgenic worms was performed 6 h after the exposure to 7.5 mM t-BOOH. (d) Intestinal localization of TRX-1 labeled with GFP. All procedures were performed as described in (a). (e) Quantitative analyses of the fluorescent images of TRX-1::GFP. All procedures were performed as described in (c) (n = 30 for each group). (f) Quantitative PCR analyses of trx-1 (n = 3). All procedures were performed as described in (c). Data are shown as the mean ± SEM. n.s.: not significant, * p < 0.05, ** p < 0.01. All p values were calculated by two-way ANOVA followed by Tukey’s multiple comparison tests.