Fig. 7

Knockdown of InR by feeding the larvae with bacterial-expressed double-stranded RNA (dsRNA). a Schematic diagram of the bacterial vector (L4440) construction. The target gene fragment was inserted in the multiple-cloning site (Xba I/Xho I) between two T7 promoter regions. The recombinant L4440 plasmids were then transformed into the RNase III-deficient E. coli strain HT115 (DE3). IPTG induces dsRNA transcription mediated by T7 promoter. b The bacteria producing InR dsRNA (dsInR) were fed to mosquito larvae (3rd instar) for 48 h; the transcription level of InR was analyzed by RT-qPCR. EGFP dsRNA-fed larvae (dsEGFP) were taken as the dsRNA control, and the larvae fed with no bacterial LB mixture were taken as the negative control (NC). c Total and phosphorylation of ERK and AKT proteins were detected by Western blotting analysis. d Gray value analysis of phosphorylated-ERK/actin (P-ERK/actin). e Gray value analysis of phosphorylated-AKT/actin (P-AKT/actin). Data are presented as mean ± SEM from three independent experiments; ns represents no significance, **P < 0.01