Figure 3: Expression analyses of DVSSJ proteins and dvssj1 mRNA.

(a) Western blot detection of DVSSJ1 and DVSSJ2 from 3^rd instar WCR dissected gut tissues. Loaded samples represent the equivalent of 1 WCR midgut. Detection of DVSSJ1 is by anti-DVSSJ1 monoclonal peptide antibody (Peptides sequence: TWNLNEEKNPDAEI 41-53 a.a.). DVSSJ2 was detected by Hybridoma Supernatant from peptide antibody production. (Peptide Sequence: MTSDTAPPDTDQRG 108-121 a.a.). The DVSSJ1 and DVSSJ2 detectable protein sizes are compared by Precision Plus Protein Western Standard (Std, Bio-Rad) ranging from 10–250 kDa (Supplementary Fig. 2). (b) Relative gene expression of dvssj1 over time by dsRNA treatment. qRT-PCR was used to examine gene expression of dvssj1. The expression represented relative expression for time points 12 and 48 hrs for treatments of double stranded RNA for dvssj1 and gus. H₂0 was used as a control treatment. Relative expression analysis was based on dvssj1 expression, after being normalized by reference gene dvrps10 expression, and then compared to dvssj1 expression in H₂0 control at each time point. Each value was shown as values ± S.E.M. of individual insects. Letter differences represent treatments that are significantly different from each other (P-value < 0.05) determined by one-way analysis of variance (ANOVA) followed by Tukey’s post-test.