Fig. 5.

Knockouts and RNAi knockdown confirm the role of Fas2 in evolved differences in pupation site choice. (A) The pupation indices of the Fas2^eb112 and Fas2^G0293gene disruptions are shown for the comparison between the original hybrid cross (Lhr; Fas2^eb112 N = 50, Fas2^G0293 N = 53) and a cross to the Drosophila melanogaster T.4 strain (Fas2^eb112 N = 51, Fas2^G0293 N = 52). Also shown for Fas2^eb112is the comparison for crosses to the D. simulans strain Mex180 (N = 53) and the D. melanogaster strain T.1 (N = 52). Asterisks denote significance (Wilcoxon tests; **P < 0.01, *** = P < 0.001). (B) A quantitative complementation interaction plot showing the proportion of females that pupated on the food when the Fas2 mutant allele (gray, circle) and the balancer chromosome (black, square) were present in offspring resulting from the D. melanogaster and D. simulans crosses shown in (A). Plotted are the least squares means from the analysis in supplementary table S5C, Supplementary Material online. The interaction was significant for Fas2 (ANOVA: P < 0.0001, determined using arcsine transformed data). (C) The results of panneuronal knockdown of the Fas2 transcript via RNAi. The proportion of RNAi or control individuals that pupated on the food are shown for the control cross (elav-Gal4 driver crossed to the RNAi background stock; N = 48), the Gal4-1 hairpin RNA cross (Gal4-1; N = 43), and Fas2 RNAi cross (N = 42). Asterisks denote significance based on Wilcoxon tests after correcting for multiple comparisons (****P < 0.0001).