HR3-knockdown <i>Aedes</i> mosquitoes showed a delay in impaired vitellogenin (Vg) mRNA and Vg protein levels at the end of vitellogenesis in the fat body.

<p>(A–C) Female mosquitoes were injected with 1 µg dsHR3 or dsMal RNAi, as described in Materials and Methods. HR3 (A and B) and Vg (C) mRNA expression was measured using qPCR at the indicated time points after blood feeding (PBM). Point 0h represents 72h post-eclosion in female mosquitoes prior to blood feeding; this was used as a reference control. Data are expressed as fold induction relative to S7. Data (means <u>+</u> standard errors of the means) from three independent experiments are shown. *Indicates statistical significance <0.05. (B) Magnification scale of the 36-h and 42-h PBM time points from the same experiment shown in A to demonstrate details of the HR3 depletion at the end of the vitellogenic cycle. (C) Monitoring the Vg transcript abundance in the same experiments as in A. A high level of the Vg transcript was observed in FBs of HR3-depleted female mosquitoes at 36 h PBM showing a delayed high expression of the <i>Vg</i> gene. (D) Western blot analysis of Vg protein during several time points of the first gonadotrophic cycle in the fat body of female mosquitoes treated with dsMal or dsHR3 RNAi. A total of 0.01 fat body equivalents was loaded in each lane for SDS/PAGE. A mixture of nine monoclonal antibodies against Vg small subunit was used for Vg protein detection (upper panels). Lower panels with Comassie-stained SDS/PAGE gels used for the western blot experiment are presented as loading controls. Three independent replicates of the experiment were made.