Fig. 3

Water-loss dehydration increases ecdysone and Malpighian tubule (MT) expression of ecdysone biosynthetic genes in young females (7 days). a 20E titer (pg/fly) (mean and SEM among three biological replicates, each of 25–30 pooled females) from whole females of two genetic backgrounds (yw and wDah) with control (food or PBS) and desiccation treatments. Independent biological replicates were prepared on separate days. One-way ANOVA within each genotype, Tukey’s post hoc comparison: *p < 0.05, **p < 0.01. b Relative disembodied (dib) mRNA in female tissues from control (food or PBS) and desiccation treatments. All values normalized to mean value from food treatment, head sample; mean values of three biological replicates, each containing the specific tissue from 15 females. Statistical comparisons are made within each tissue by t test for difference between desiccation and PBS treatment: dib mRNA was significantly elevated in muscle and MT (*p < 0.05 with Holm–Bonferroni Sequential correction). c Relative mRNA for EcR and (d) PGRP-LC in control genotype (+/dib(RNAi)) and when dib is depleted in stellate cells (c724>) or principal cells (c324>). Values are mean and SEM of three independent biological replicates, normalized relative to food treatment, wild-type for each mRNA type. For each genotype: one-way ANOVA with Tukey’s post hoc comparison. For the control genotype (+/dib RNAi) a significant difference was observed between the flies exposed to desiccation compared to those exposed to normal food or PBS (*p < 0.05, **p < 0.01). However, no significant differences were observed when EcR or PGRP-LC were knocked down (c324 > RNAi and c724 > RNAi); EcR-RNAi F = 0.52, p = 0.62 and F = 4.43, p = 0.066; PGRP-LC-RNAi F = 0.39, p = 0.70 and F = 0.54, p = 0.61