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. 2009 May 19;86(3):633–643. doi: 10.1189/jlb.1008662

Figure 3.

Figure 3.

Ethyl pyruvate modifies intestinal RAGE expression. Paraformaldehyde-fixed colons of (A, C) wild-type C57BL/6, (D) IL-10−/− mice, or (E) IL-10−/− mice treated with ethyl pyruvate were stained for actin (red) and RAGE (green) and viewed by confocal microscopy. (B) Absence of immunostaining of the wild-type colon with an isotype control antibody (nuclei-stained blue with Draq5). Colocalization of actin and RAGE is represented by a yellow color observed at the luminally oriented microvillus border (white arrows in C and E). Each image is a representative result from at least five different mice. Immunoflourescence demonstrates down-regulation of RAGE in the colonic epithelium of IL-10−/− mice compared with wild-type controls (C vs. D). In ethyl pyruvate-treated IL-10−/− mice (E), RAGE staining appears similar to wild-type colon. (F) Fecal extracts isolated from wild-type (n=5), IL-10−/− mice treated with vehicle (n=4), or IL-10−/− mice treated with ethyl pyruvate (ethyl pyruvate, n=4) were analyzed for RAGE by ELISA. Levels were normalized to total fecal protein. Each result represents the mean ± sd (P=0.05 for wild-type vs. IL-10, vehicle-treated; P=0.06 for wild-type vs. IL-10, ethyl pyruvate-treated). (G) LPMCs from wild-type (WT; upper panel, left) and IL-10−/− (upper panel, right) mice were stained with RAGE and F4/80 (macrophage marker) and analyzed by flow cytometry. As a control, splenocytes from wild-type (lower panel, left) and IL-10−/− (lower panel, right) mice were stained with RAGE and F4/80 for flow cytometry. Results are representative of three experiments with similar results.