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. Author manuscript; available in PMC: 2014 Feb 1.
Published in final edited form as: Hypertension. 2012 Nov 26;61(2):443–449. doi: 10.1161/HYPERTENSIONAHA.112.196303

Figure 1.

Figure 1

Characterization of long-term rat inner medullary cells (A-I). IM cells show specific immunoexpression of AQP-2 (red; A), AT1R (red; B), COX-2 (green; C), and PRR (green; D). AT1R (red) co-localizes with COX-2 (green) in the same type of cells (E), indicating that interstitial cells (arrows) strongly stained for COX-2 (green) also co-express AT1R (red) in the plasma membrane, as previously described (E). Cells expressing AQP-2 (red; F), the principal cells, do not co-express COX-2 (arrow, green; F). Tenascin C (arrow, green; G), a marker for interstitial cells, does not co-localize with AQP-2 (red; E). Likewise, anion exchanger type-1 (AE-1; red; H), a known immunomarker for intercalated type-A cells (arrowheads), does not co-localizes with AQP-2 (green; H). Evidence for the presence of PRR (red; I) co-localizing with COX-2 (arrow heads, green; I) is shown in I. Nuclei were counterstained with DAPI (4′,6-diamidino-2-phenylindole; blue). In addition, in normal rat kidney sections (3 μm) immunofluorescence studies demonstrate that PRR (apical green immunoreactivity) and AE-1 (basolateral red immunoreactivity) co-localize in tubular intercalated type-A cells (J). K shows specific immunoreactivity for COX-2 in the interstitial cells (arrows) and some tubular cells (arrowheads) in the rat inner renal medulla (brown chromogen, 3,3′-Diaminobenzidine DAB). In consecutive kidney sections (L and M), is evident that PRR (green; L) is immunoexpressed by negative AQP-2-expressing cells (red; M). Panel N displays examples of the co-localization of PRR (red; cell membrane) with COX-2 (green; intracellular localization) in the intercalated cells of the collecting ducts. In addition, this panel contains a high resolution microphotograph (left upper corner, 100X, oil immersion) for clear details. The lower panels O–Q show microphotographs (20X magnification) of a rat kidney section stained using dual immunofluorescence for COX-2 (red; O) and PRR (green; P) counterstained with DAPI (blue fluorochrome) demonstrate that COX-2 and PRR also co-localized in interstitial cells (merge, Q). Images were visualized using a Nikon Eclipse 50i immunofluorescence microscope and microphotographs were captured using a digital camera Nikon DS-U2/L2.