Figure 2. Oral infection with a HCD quorum-sensing-defective V. cholerae mutant leads to decreased activation of ecdysone-responsive, innate immune, and epithelial repair genes.

(A and B) Gene Ontology (GO) analysis and (B) volcano plot of significantly regulated genes in the fly intestine during oral infection with WT V. cholerae compared with a ΔhapR mutant. Analysis is based on an RNA-seq experiment with biological triplicates that included an expression fold change greater than 2 with adjusted p < 0.05 to be significant. The calculated false discovery rate (FDR) is shown.

(C and D) qRT-PCR analysis of (C) the indicated 20E-regulated genes and (D) transcription of the IMD pathway-regulated AMPs DptA, CecA1, and AttA in the intestines of OreR flies orally infected with WT V. cholerae or a ΔhapR mutant.

(E and F) qRT-PCR transcriptional analysis of (E) the V. cholerae HapR-regulated genes hapA and vpsL and (F) the indicated IMD pathway-regulated AMPs in the intestines of Tk > driver only and Tk > Tk^RNAi flies orally infected with WT V. cholerae or a ΔhapR mutant. Measurements for ΔhapR-infected flies were normalized to that of WT V. cholerae-infected flies of similar genotype. For qRT-PCR results, the mean of biological triplicates is shown. Error bars represent the standard deviation. A Student’s t test was used to assess significance.

(G) Fractional survival over time of isogenic control and mutant flies with deletions in genes encoding the AMPs Def, AttC, Dro, AttA, AttB, DptA, Drs, and AttD fed WT V. cholerae or a ΔhapR mutant.

Significance was calculated by log-rank analysis. **p < 0.01, *p < 0.05, **p < 0.01, ***p < 0.001. See also Figures S2 and S3 and Table S1.