Fig. 7. Immunolocalization of AgNHA1 (green; FITC) and P-type Na⁺/K⁺ ATPase (red; TRITC) counterstained with a nuclear DNA marker (pseudo-blue; DRAQ-5) in isolated tissues of fourth instar An. gambiae larvae. (A) A composite whole mount, showing AgNHA1 expressed in the gastric caeca (GC), anterior midgut (AMG), selected cells of the posterior midgut (PMG), proximal Malpighian tubule (MT) and in a subset of cells in the rectum. (B) AgNHA1 was expressed in two bands of highly immunoreactive cells in the cardia. (C) AgNHA1 was highly expressed in the caudal tip of the gastric caeca. Scale in C is the same as B. (D) In the anterior midgut, AgNHA1 expression appeared to have a vesicular pattern of staining. (E) AgNHA1 stained cells (arrows) in the PMG and intensely stained cells in the proximal portion of the MTs (see arrow). Scale in E is the same as D. (F) AgNHA1 is expressed in the subset of cells in the rectum. AgNHA1 staining did not occur in the dorsal anterior rectal (DAR) cells. Similarly, Na⁺/K⁺-ATPase stained much of the rectum but did not stain its DAR cells (G). (H) An overlay of the AgNHA1 and Na⁺/K⁺-ATPase expression in the rectum. Colocalization of AgNHA1 and Na⁺/K⁺-ATPase in the same cells is indicated by the orange staining pattern. Scale in G,H is the same as F. In addition, AgNHA1 was expressed in the neuropil of the sub-esophageal ganglion (SOG) (I) and the abdominal ganglion (AG) (J). The scale in J is the same as in I.