Fig. 4

Reproductive phenotypes associated with selected RNAi in response to dsRNA injection immediately after blood feeding. A Schematic diagram of experimental time course for dsRNA microinjection, blood feeding, and oviposition in the first gonotrophic cycles. Mosquitoes were microinjected with dsRNA immediately after blood feeding. B Representative eggs shown are from four different mosquitoes microinjected with either dsRNA-Fluc, -Nasrat, -Closca, -Polehole, or -Nudel. Female mosquitoes microinjected with dsRNA-Nasrat, -Closca, or -Polehole showed no difference in egg production (C), melanization (D), and embryo viability (E) compared to RNAi-Fluc control mosquitoes. Females microinjected with dsRNA-Nudel immediately after blood feeding produced the similar number of eggs as controls, but had greatly reduced eggshell melanization and viability. The effect of selected RNAi on A. aegypti egg production was examined by counting the number of eggs laid by each individual female. Each dot represents the number of eggs oviposited by an individual mosquito (C). Egg melanization was examined under a light microscope (D) and viability was determined by hatching them 1 week after oviposition (E). Each bar corresponds to egg viability from 12 individual mosquitoes from three groups. 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 6: Table S5