Aedes aegypti (L.) (Rockefeller strain) mosquitoes were used for all studies. Larvae were maintained on a diet consisting of equal proportions of rat chow (Sunburst Pet Foods, Phoenix, AZ), lactalbumin hydrolysate (USB, Cleveland, OH), and yeast hydrolysate (USB, Cleveland, OH). Female pupae were separated Figure 8 . Quantitation of whorl size and number in dsRNA injected unfed and amino acid fed mosquitoes. Whorls with a minimum of five stacked ER membranes were counted and the total area of each whorl was determined as described in figure 2 legend. A) Size and number of RER whorls in 20 random fields from unfed and amino acid fed mosquitoes after 120 min. that were injected with Fluc or alpha-COPI dsRNA. B) Total whorl in the same sample as shown in A. Statistical analysis using Pearson Chi-square test revealed that the total whorl area in alpha-COPI dsRNA injected unfed and amino acid fed mosquitoes was significantly less than the total whorl area in Fluc dsRNA injected unfed mosquitoes (bars with different letters signify significance at p,0.001). In addition, the total whorl area in Fluc dsRNA injected fed mosquitoes was found to be significantly less than in Fluc dsRNA injected unfed mosquitoes using the Pearson Chi-square test (bars with different letters signify significance at p,0.001). doi:10.1371/journal.pone.0018150.g008 from males using a mosquito separator. Adult mosquitoes were routinely maintained at 28uC, 70-80% relative humidity and a photoperiod of 16:8 h (L:D), on 10% sucrose ad libitum.