Fig. 7

Reproductive phenotypes following RNAi knockdown of DCE2, DCE4, DCE5, and CATL3 in Aedes aegypti mosquitoes. A Representative eggs are shown from mosquitoes microinjected with dsRNA against Fluc control, DCE2, DCE4, DCE5, and CATL3. B An effect of RNAi on fecundity was studied during the first gonotrophic cycle. Each dot represents the number of eggs oviposited by an individual mosquito. C Melanization of these eggs was examined under a light microscope and a melanization percentage was determined. D An effect of RNAi on viability of eggs was examined by hatching eggs 1 week after oviposition. Each bar corresponds to egg viability from 23 to 26 individual mosquitoes from three independent cohorts. The mean ± SEM are shown as horizontal lines. Statistical significance is represented by stars above each column (unpaired Student’s t test; *** P < 0.001, NS not significant). A detailed phenotypic analysis is shown in Additional file 10: Table S9. RNAi-mediated knockdown efficiency was validated by quantitative real-time PCR (qPCR). Relative abundance of mRNA levels for DCE2 (E), DCE4 (F), DCE5 (G), and CATL3 (H) was analyzed in dissected mosquito ovaries at 36 h PBM. Mosquitoes were microinjected with each dsRNA at 4 days prior to blood feeding, as shown in Fig. 1. dsRNA-Fluc-injected mosquitoes were used as controls. A single mosquito analysis was performed to isolate total RNA, synthesize cDNA, and monitor silencing efficiency by qPCR. mRNA levels were normalized according to transcript levels of ribosomal S7 protein. Data are presented as MEAN ± SEM of 12 individual mosquitoes. *** P < 0.001 compared to RNAi-Fluc