Figure 1. Genetic modulation of dRYBP expression levels affects AMP production.

(A, B, C, D, E). (A) All tested dRYBP mutants show an excessive immune response to Gram-negative infection. Adult females of indicated genotypes were left unchallenged (left panel) or infected by pricking with Ecc15 (heat-killed at 95°C during 15 min) and collected at 8 h (middle panel) and 24 h (right panel) after infection for Diptericin (Dpt) mRNA quantification. The complete data set (unchallenged, 8 h, 24 h) was analysed by two-way ANOVA with Bonferroni post-tests, using Can^S as control. (B) dRYBP mutants also show excessive Imd activation after infection with another Gram-negative pathogen, E. coli. Adult females of indicated genotypes were treated as in (A). (C) Infection time-course in wild-type, heterozygous and homozygous dRYBP^Δ55 mutants. Flies were treated as in (A). (D) Loss of dRYBP affects at least two Imd-dependent antimicrobial peptides. mRNA levels of Attacin-B (AttB) were quantified after infection as described in (A). (E) Overexpression of dRYBP reduces the Imd response to Gram-negative infection. Adult females were infected as in (A), then were heat-shocked for 1 h at 37°C and collected at 8 h and 24 h after infection for quantification of Dpt mRNA levels. (F) dRYBP expression levels are not affected by infection. Wild-type adult females were infected as in (A) and dRYBP mRNA levels were monitored over time. For all graphs, data represent mean + SEM of at least 3 biological repeats, and asterisks denote the following p values: *, 0.01<p<0.05; **, 0.001<p<0.01; *** p<0.001; ns, not significant.