The PLC/Ca2+/CaMKII pathway is essential for the JH-induced gene expression in Ae. aegypti. (A) Fat bodies from newly emerged mosquitoes were cultured in vitro with the indicated inhibitors for 1 h. After JH-III was added to the culture medium, the fat bodies were cultured for another hour and collected for RNA extraction. Expression of the JH target genes AaKr-h1 and AAEL002576 was analyzed by real-time PCR. Results are the mean ± SD of three independent experiments. (B) Female mosquitoes at 1 h PE were injected with 0.5 µg of dsRNA against GFP (a negative control), individual PLC isoforms, CaMKI, or CaMKII. At 72 h after the injection, the mosquitoes were collected for RNA extraction. Expression of the JH-regulated genes in adult mosquitoes was measured using real-time PCR. Results are the mean ± SD of three independent experiments.