Figure 8

Confirmation of developmental stage- and sex-specific expression of SP and SPH genes by qRT-PCR. Total RNA was extracted from eggs, 2nd instar nymphs, 5th instar nymphs, female adults and male adults, individually, and used for the expression analysis of SP/SPH genes using qRT-PCR. The relative expression levels of each gene in each developmental stage or sex were normalized using the N. lugens 18 s rRNA Ct values. Three biological replications were conducted and the ΔΔCt method was used to measure the relative transcript levels in each developmental stage. Results of triplicate experiments are shown with the standard deviations. The asterisk (**) indicates statistical significance at p < 0.01. 2nd, 5th, F and M refer to the 2nd instar nymphs, 5th instar nymphs and female and male adults, respectively.