Fig. 1: Characterization of C. quinquefasciatus salivary long D7 proteins.

a Gene expression analysis of CxD7L1 and CxD7L2 transcripts in different stages of C. quinquefasciatus mosquitoes. Relative abundance was expressed as the fold change using the 40S ribosomal protein S7 as the housekeeping gene. Larvae stage 1 (L1), larvae stage 2 (L2), larvae stage 3 (L3), larvae stage 4 (L4), pupae, male adult (reference sample), female adult, heads and thoraxes (H+T), and abdomens from female adult mosquitoes were analyzed separately. Two biological replicates and two technical duplicates were analyzed. Bars indicate the standard error of the means. b Purification of CxD7L1 (blue line) and CxD7L2 (red line) by size exclusion chromatography using Superdex 200 Increase 10/300 GL column. c Coomassie-stained NuPAGE Novex 4–12% Bis–Tris gel electrophoresis (n = 1) of recombinant proteins CxD7L1 and CxD7L2 (1.5 µg). SeeBlue Plus2 Pre-stained was used as the protein standard (M). d, e Immunolocalization of CxD7L1 and CxD7L2 proteins in the salivary glands of C. quinquefasciatus. Salivary glands were incubated with rabbit IgG anti-CxD7L1 (d), anti-CxD7L2 (e), and further stained with anti-rabbit IgG Alexa Fluor 594 antibody shown in red. Proteins of interest were localized in the medial and distal regions of the lateral lobes of C. quinquefasciatus salivary glands. As a control, salivary glands were incubated with anti-rabbit IgG AF594 alone (f). Nucleic acids were stained by DAPI (blue) and the actin structure of salivary glands was stained using Phalloidin Alexa 488 (green). Four independent experiments were performed with 1–2 glands imaged per experimental group. Scale bar = 50 µm. Source data are provided as a Source Data file.