Fig. 2

Duct appearance differs outside versus inside of the salivary glands. a DIC imaging of DAPI-stained An. stephensi female SGs highlighting the duct. Ladder-like taenidia are obvious in the portions of the duct proximal to the SG (Ai-ii, iv-v). Once inside the gland, duct morphology becomes smooth (Aii-iv, vi), continuing all the way through the termini (Aiii). b-d Confocal images of adult An. stephensi SG lobes stained with rhodamine-conjugated wheat germ agglutinin (WGA; chitin, O-GlcNAc groups) and DAPI (nuclei). (2Bi) Maximum intensity projection of WGA signal (greyscale) throughout an entire female SG as well as the individual duct (ID) and part of the common duct (CD) is shown. The triductal junction (TDJ) within the SG is highlighted (red) in the zoom image shown in Bii. Duct chitin levels are higher inside the SG (yellow arrow) than outside (white arrow). Biii (90^o counter-clockwise rotation of boxed region in Bi) shows one ID as it branches away from the CD. Biv is a zoom of the boxed region in Biii, highlighting SG duct organization. In the CD and ID, cells/nuclei are typically found in close association with the duct. A low percentage of individual CD and ID cells/nuclei are more tightly associated with the basement membrane (arrow in Biv). c Confocal images of female SGs stained with WGA and DAPI (nuclei). Rare nuclei are observed in close proximity to the duct (Dviii, yellow arrows). d Confocal images of male SGs stained with WGA and DAPI. WGA staining labels the chitinous salivary duct and O-GlcNAcylated proteins within SG cells. Five fused duct termini are marked by asterisks (Diii). Scale bar lengths are: 50 microns—Bi, Ci, Civ, Cvii, Di; 20 microns—Ai, Aiii, Aiv, Bii, Biii, Ciii, Cvi, Cviii, Diii, Div; 10 microns—Av, Biv; 5 microns: Avi. CD: common duct, ID: individual duct, SG: salivary gland, SD: salivary duct (in gland), SDT: salivary duct terminus, TDJ: triductal junction