Fig. 2

RNAi knockdown efficiency and chymotrypsin-like activity determination in individual Ae. aegypti mosquito midgut tissue extracts. Representative western blot (WB) of mosquito midgut extracts at 3 h PBM (A) and 18 h PBM (B). α-tubulin was used as an internal control. The WBs in (A and B) were cropped for clarity. The original blots are presented in Supplementary File 1. Quantification of AaCHYMO protein expression (C) and quantification of mRNA levels via qPCR (D). Data was obtained from individual midgut of 12 mosquitoes. Data are presented as mean ± SEM. Statistical significance is represented by stars above each comparison (unpaired Student’s t-test; **** p < 0.0001 compared to the RNAi-FLUC control samples). Detection of chymotrypsin-like activity in FLUC vs. RNAi AaCHYMO mosquito midgut extracts using the MeO-Suc-Arg-Pro-Tyr-AMC substrate (E). Activity is eliminated in the 3 h to 24 h PBF samples, with an overall significant reduction in chymotrypsin-like activity at 36 h PBM (****p < 0.0001) and at 48 h PBM (**p < 0.05).