Figure Legends
Figure 1. Overall effects of organophosphate (OP) and pyrethroid (Pyr) factors on survival and development. Hazard ratios depicting the effects of selection regime (OP Reg or Pyr Reg), pesticide exposure (OP PTx or Pyr PTx), Bt treatment (BtTx), the interaction between selection regime and pesticide exposure (OP Reg:PTx or Pyr Reg:PTx), the interaction between selection regime and Bt treatment (OP Reg:BtTx or Pyr Reg:BtTx), and the interaction between pesticide exposure and Bt treatment (OP PTx:BtTx or Pyr PTx:BtTx) on survival (A, B) and development (C, D) are shown in the left panels. Raw data illustrating the proportion of surviving individuals (A, B) or the proportion of adults (C, D) are shown in the right panels. For hazard ratio plots depicting survival and development estimates, positive significant effects (higher survival or faster development) are colored blue, negative significant effects (lower survival or slower development) are red, and nonsignificant effects are black (Suppl. Table 2).
Figure 2. The influence of pesticide exposure on septic infection outcomes: control-regime larvae exposed to OP are more susceptible to Btt septic infection. The proportion of surviving individuals 24h post septic Btt infection are shown for control (Regime –) and OP selection regime (Regime OP) larvae exposed to control (Pesticide Tx –) or OP diet (Pesticide Tx OP) treatments. Circles indicate surviving proportions of individuals per block. Significant differences between groups are denoted with asterisks (p < 0.0005; Suppl. Table 6).
Figure 3. Number of significantly differentially expressed (sig. DE) genes (padj < 0.05) for factors within each DESeq2 model (number of sig. DE transcripts are denoted above bars; relative to control regime or no pesticide/ Bt treatments, positive counts are upregulated transcripts and negative counts are downregulated). A. No pesticide model: main and interaction effects of pesticide selection regime and Bt treatment in the absence of pesticide exposure. Hatched bars indicate no pesticide treatment in the model. B. OP model: main and interaction effects of OP selection regime, pesticide exposure, and Bt treatment. C. Pyr model: main and interaction effects of Pyr selection regime, pesticide exposure, and Bt treatment. The table below indicates the differentially expressed factor. Main effect of selection regimes = Regime OP/ Pyr. Main effect of Bt treatment = Bt Tx +. Main effect of pesticide treatments = Pesticide Tx OP/ Pyr. Interactions between selection regimes and pesticide exposure = Regime OP/ Pyr & Pesticide Tx OP/ Pyr. Interactions between selection regimes and Bt exposure = Regime OP/ Pyr & Bt Tx +. Interactions between pesticide and Bt exposure = Pesticide Tx OP/ Pyr & Bt Tx +.
Figure 4. Profiles of differentially expressed transcripts demonstrate opposing responses in Bt-infected individuals that were or were not also exposed to or selected with pesticides. Transcripts significantly differentially expressed (padj < 0.05) with Bt treatment and interactions with A. OP exposure, and B.Pyr selection regime and exposure. The DE Factor/ Model legend refers to the factor corresponding to the differentially expressed gene set and the DESeq2 model used (no pesticide- hatched bars; OP or Pyr- solid bars; BtTx = main effect of Btt treatment; OP PTx = main effect of OP exposure; OP or Pyr PTx:BtTx = interactions between OP or Pyr exposure and Btt treatment; Pyr Reg:BtTx: interaction between Pyr selection regime and Btt treatment). Colored circles on the left indicate transcripts included in the magenta WGCNA module (Suppl. Fig. 7). Transcript abbreviations are as follows: Stc = shuttle craft-like protein, Def = defensin, Att = attacin, PR = pathogenesis-related protein, HRG = histidine-rich glycoprotein, SP = serine protease, P450 = cytochrome P450, CEH = carboxylic ester hydrolase, GST = glutathione S-transferase, ABC = ABC transporter, Chit = chitinase protein, CP = cuticle-related protein, PM = peritrophic matrix related protein, Vit = vitellogenin, Ins = insulin-like peptide, LSP = larval serum protein, AchE = acetylcholinesterase, KLF = Krüppel-like factor, ApoIII = apolipophorin-III, Neur = neuropeptide.
Figure 5. Normalized gene counts illustrate dampening effects of Pyr selection regime and OP treatment on immune gene expression induced with Bt expsoure. A. OP (OP model- OP PTx) and Bt treatment (OP model- BtTx) effects are shown for TC007737 (attacin1), and B. Pyr Regime (Pyr model- Pyr Reg) and Bt treatment (Pyr model- BtTx) effects are shown for TC010517 (defensin-like).