Characterization of the ability of AgNimB2 and AgEater to mediate bacterial phagocytosis. (A) RNA interference (RNAi) gene knockdown (KD) efficiency compared to S7 reference from 20 of each dsRNA injected mosquitoes assayed by quantitative real-time PCR (qRT-PCR). dsAgNim KD efficiency, 99.2±0.02%; dsEater KD efficiency, 99.8±0%. qRT-PCR data analyzed using standard curve method. Error bars represent standard error of the mean for two replicates. (B) Fluorescent images from pHrodo bioparticle injected mosquito abdomens, example of phagocytosis foci are indicated by arrows. (C) pHrodo phagocytosis foci counts from AgNimB2 KD mosquitoes challenged with Staphylococcus aureus pHrodo bioparticles, (D) pHrodo phagocytosis foci counts from AgNimB2 KD mosquitoes challenged with Escherichia coli pHrodo bioparticles, (E) pHrodo phagocytosis foci counts from AgEater KD mosquitoes challenged with E. coli pHrodo bioparticles, (F) pHrodo phagocytosis foci counts from AgEater KD mosquitoes challenged with S. aureus pHrodo bioparticles. Each dot represents one mosquito, with three replicates shown for each gene KD. Bar plot indicates median (bold line), inter quartile range (IQR), and 1.5×IQR (dotted line). Medians were compared using a Wilcoxon rank sum test with continuity correction.