Female salivary gland–specific expression of AAPP. (A) Western blotting for AAPP in the salivary glands of female mosquitoes. A homogenate of 2 pairs of female salivary glands was subjected to 10% SDS-PAGE. Lane 1, silver staining; lane 2, Western blotting using a mouse anti-rAAPP immune serum. (B) RT-qPCR of aapp mRNA. Total RNA was isolated from salivary glands (SG) and carcasses (Ca) of male and female mosquitoes. The expression of aapp mRNA was examined for its tissue- and sex-specificity using RT-qPCR. The fold induction was determined from threshold cycle values normalized for ubiquitin mRNA expression and then normalized to the values of each gene from the male salivary gland mRNA. (C) Immunostaining of salivary glands with the anti-rAAPP immune serum. The paired salivary glands of mosquitoes are present in the thorax flanking the esophagus. Each gland consists of 3 lobes: a distal lateral lobe (DL), proximal lateral lobe (PL), and medial lobe (M). (i) phase-contrast image of the salivary glands of a wild-type mosquito; (ii) immunostaining of the salivary glands of a wild-type mosquito with the anti-rAAPP immune serum; (iii) phase-contrast image of the salivary glands of a DsRed transgenic mosquito; and (iv) DsRed expression in the salivary glands of a DsRed transgenic mosquito. Scale bars: 100 μm.