Fig. 4.

Blocking the formation of AaTai-A and AaTai-B by the knockdown of AaSRPK or AaClk arrests ovarian development. DsRNAs for AaSRPK and AaClk were injected into female mosquitoes within 30 min PE. DsRNA for the GFP gene (dsGFP) was used as a control. The knockdown of AaSRPK and AaClk was validated at 96 h PE (A) and 24 h PBM (B) using qRT-PCR. The results are presented as the mean ± SD of three independent experiments (**P < 0.01). The expression of AaTai isoforms was examined using RT-PCR at 96 h PE (C) and 24 h PBM (D). Noninj, noninjected. (E and F) Images of representative ovaries that were isolated from the Noninj and dsRNA-injected female mosquitoes at 96 h PE and 24 h PBM. (Scale bars: 500 µm.) (G and H) Average lengths of primary follicles at 96 h PE and 24 h PBM. The sizes were measured using the Leica Application Suite (v4.5). Thirty individuals in each group were used, and the results are shown as the mean ± SD. (I) Expression of the JH response genes AaKr-h1 and AAEL002576 was examined by qRT-PCR in the fat body at 96 h PE. qRT-PCR results are presented as a fold change compared with the dsGFP-injected sample (P > 0.05). (J) Expression of the 20E response genes AaE74B, AaE75A, and AaVg was measured by qRT-PCR at 24 h PBM. Statistical significance is shown (**P < 0.01).