Figure 2. Impact on ecdysial behaviors of manipulating ETH effectiveness on downstream targets.
(A) Ecdysial behaviors in intact puparium. Duration of pre-ecdysis (left; open bars) and ecdysis (right; filled bars) in controls (CS and CCAP>GCaMP), and in animals expressing ETHR RNAi in different subsets of ETH targets. (B) Corresponding ecdysial behaviors of puparium-free preparations. Data are mean ± SEM. In (A) and (B) duration of ecdysial phases is indicated as not significantly different (‘ns') or significantly different ('*': p<0.5; '**’: p<0.01) than those of CCAP>GCaMP control (one-way ANOVA, Dunnett's post-hoc to control). Comparison of frequency of contractions per minute for pre-ecdysis and ecdysis (labeled Fpre and Fecd, respectively) in puparium-free preparations is indicated as not significantly different (‘ns') or significantly smaller ('<': p<0.05; '<<’ p<001) or greater ('>': p<0.05; '>>’ p<001) than that of CCAP>GCaMP controls (one-way ANOVA, Dunnett's post-hoc to control). Actual p values can be found in Supplementary file 1. Genotypes: all animals expressed GCaMP under control of Ccap-GAL4 (Ccap-GAL4 + UAS-GCaMP). ETHR RNAi: UAS-ETHR RNAi; Df(ETHR)/+: hemizygosity for ETHR; MN>: motoneuron GAL4 (C164; see Materials and methods). In all experiments using RNAi, its effectiveness was boosted by including a UAS-dcr2 transgene.