FIGURE 8.

Changes in HS sulfation in murine model of fibrosis. UUO mice were generated. On day 7 the animals were sacrificed, and the kidneys were retrieved. A, frozen section of UUO and unoperated control kidneys were stained with antibodies against collagen-I and HS (HS3A8 and RB4EA12), respectively. Sections stained with collagen antibody (1:50) were visualized by incubating with biotinylated secondary antibody, and color was developed with 3,3-diaminobenzidine solution (brown). HS sulfation was examined using phage display antibodies HS3A8 and RB4EA12 followed by staining with Cy3-conjugated anti-VSV antibody. B, quantitative analysis was performed by measuring the density of tubular and interstitial areas randomly by Image J software (3–5 areas/kidney). Three animals were investigated. ***, p < 0.001; **, p < 0.01.