(A) Impact of Kayak depletion on expression of genes related to complement system (TEP20, TEP24, TEP15 and TEP2), apoptosis (Caspase8 and Dronc) and autophagy (ATG14 and ATG18A) at 10 days post ZIKV oral infection. Gene expression was quantified in pools of 10 salivary glands. Actin expression was used for normalization. Bars show arithmetic means ± s.e.m. from three biological replicates. (B-D) Effect of depletion of (B) Dronc or TEP20 alone, (C) Dronc and TEP20 simultaneously, and (D) Puc alone, or Puc and Dronc simultaneously, or Puc and TEP20 simultaneously on salivary gland infection at 7 days post ZIKV inoculation. Four days prior infection, mosquitoes were injected with dsRNA. Bars show geometric means ± 95% C.I. from 20 individual salivary glands. Each dot represents one sample. TEP, Thioester-containing protein; ATG, Autophagy related gene; dsKay, dsRNA against Kayak; dsPuc, dsRNA against Puckered; dsCtrl, dsRNA against LacZ; dsTEP20, dsRNA against TEP20; dsDronc, dsRNA against Dronc. *, p < 0.05; **, p < 0.01; ***, p < 0.001, as determined by unpaired t-test (A) or Dunnett’s test with dsCtrl as control (B-D).