Fig 2. A single microinjection of dsRNA against EOF1 promotes the abnormal eggshell phenotypes in multiple gonotrophic cycles.
(A) Schematic diagram of experimental time course for dsRNA microinjection, blood feeding, and oviposition in the first three gonotrophic cycles. Representative eggs are shown from mosquitoes microinjected with dsRNA-Fluc (B) and dsRNA-EOF1 (C). An effect of RNAi-EOF1 or RNAi-Fluc control on fecundity was studied during the first three gonotrophic cycles (D). Each dot represents the number of eggs oviposited by an individual mosquito (N = 30). The mean ± SE are shown as horizontal lines, and statistical significance is represented by stars above each column (unpaired Student's t test; ***p < 0.001; **p < 0.01). An effect of RNAi-EOF1 on viability was also examined during the first three gonotrophic cycles (E). A significant reduction in the percentage of egg hatching was observed in RNAi-EOF1 mosquitoes. Each bar corresponds to egg viability from 15 individual mosquitoes from both groups. Underlying data can be found in S1 Data. dsRNA, double-stranded RNA; EOF1, eggshell organizing factor 1; Fluc, firefly luciferase; RNAi, RNA interference; SE, standard error.