RNA interference efficiency and the effect of dsInR on ovarian follicle length in C. pipiens. (A) cInR transcript levels in females injected with dsInR were reduced compared with the dsβ-gal controls. Expression levels were measured by RT-PCR 4 and 7 days after dsInR (RNAi) injection, using dsβ-gal as a control and expression of 28S rRNA (30 cycles) as a loading control. (B) Effect of dsInR knockdown on ovarian development of females programmed for nondiapause. Injections of dsInR (≈0.7 μg per female) were made into the thorax of cold-anesthetized mosquitoes within 1 day after eclosion (controls, dsβ-gal), and females were dissected 10 days later. ND, programmed by long day length for nondiapause; D, programmed by short day length for diapause. Both groups were maintained at 18°C. Bars (mean ± SD) with the same letters are not significantly different at P = 0.05, ANOVA.