Supplemental Figure Legends
Suppl. Figure 1. Experimental design schematic. Larvae from six distinct populations were exposed to control, organophosphate (OP), or pyrethroid (Pyr) diets for at least eight generations. Adults from Gen. 8 pesticide-selected and control populations were allowed to reproduce on control diets, and twelve-day old F2 larvae from each population were exposed to control and pesticide diets with and without Btt .
Suppl. Figure 2. In vitro growth rate (r) of Bttunder control and pesticide conditions. A. In vitrogrowth rate (r) of Btt under control and OP conditions. Data were not normally distributed, so non-parametric tests were used. There was a significant difference between replicates, so the effect of treatment was analyzed for each replicate separately (Suppl. Table 4). K-W test revealed significant positive effect of OP on Btt growth rate for three of the four replicates (reps 2-4; Suppl. Table 4). B.In vitro growth rate (r) of Btt under control and Pyr conditions. Data were not normally distributed, so non-parametric tests were used. There was no significant difference between replicates or with the effect of treatment (Suppl. Table 4).
Suppl. Fig. 3. Germination of spores from diet disks in the absence or presence of either pesticide (Bt, OP + Btt or Pyr +Btt : n=10, control: n=6). There was no significant difference between Bt control and Bt pesticide diet disks (Suppl. Table 5).
Suppl. Figure 4. Top significantly enriched GO terms for differentially expressed gene sets. A. Biological process GO terms, B. Molecular function GO terms. Differentially expressed gene sets are denoted on the x-axis and refer to the DESeq2 model (no P: = no pesticide model, OP: = OP model, Pyr: = Pyr model) and the factor corresponding to the differentially expressed gene set (BtTx = main effect of Btt treatment; OP Reg, Pyr Reg = main effect of OP and Pyr selection regimes, respectively; OP PTx, Pyr PTx = main effect of OP and Pyr exposure, respectively; OPReg:PTx, PyrReg:PTx = interaction between OP and Pyr regime and exposure, respectively; PyrReg:BtTx = interaction between Pyr regime and Btt treatment; OPTx:BtTx = interaction between OP exposure and Btt treatment).
Suppl. Fig. 5. Canonical detoxification genes significantly differentially expressed (padj < 0.05) with A. OP exposure (OP PTx), OP selection regime (OP Reg), Btt exposure (BtTx), and the interaction between OP regime and exposure (OP Reg:PTx) and B. Pyr exposure (Pyr PTx), Pyr selection regime (Pyr Reg),Btt exposure (BtTx), and the interaction between Pyr regime and exposure (Pyr Reg:PTx).
Suppl. Fig. 6. Cuticle-associated genes significantly differentially expressed (padj < 0.05) with A. OP exposure (OP PTx), OP selection regime (OP Reg), Btt exposure (BtTx), the interaction between OP regime and exposure (OP Reg:PTx), and the interaction between OP and Btt exposure (OP PTx:BtTx), andB. Pyr exposure (Pyr PTx), Pyr selection regime (Pyr Reg),Btt exposure (BtTx), the interaction between Pyr regime and exposure (Pyr Reg:PTx), the interaction between Pyr regime andBtt exposure (Pyr Reg:BtTx), and the interaction between Pyr andBtt exposure (Pyr PTx:BtTx).
Suppl. Fig. 7. WGCNA gene modules and significant relationships with experimental factors. The total number of transcripts (n) and number of differentially expressed (DE) transcripts in each module are listed, and significant relationships with experimental factors are indicated with black stars. Top values within each cell indicate the correlation strengths (corresponding to the heat map colors) and bottom values in parentheses indicate p-value significance for the relationship of each module with the different experimental factors. Significant (p < 0.05) positive correlation values and red cells indicate that the transcripts within the module are commonly upregulated; negative significant correlation values and green cells indicate that the transcripts within the module are commonly downregulated.