Fig. 4.

Syndecan-4 (Synd4) expression is increased in unilateral ureteral obstruction (UUO) kidneys (quadruple staining). A: representative images of PDGF receptor-β (PDGFRβ)-, CD31-, Synd4-, and Hoechst-stained contralateral (CLK) and UUO kidney sections from wild-type (WT) and endothelial sirtuin 1-deficient (Sirt1^endo−/−) mice (n = 4). Scale bar = 10 µm. B: quantification of Synd4 fluorescence intensity per cell. A Synd4-stained image was merged with the respective Hoechst-stained image. Cells with Synd4-positive staining were counted manually. Finally, the measured total integrated density of Synd4 staining was divided by the respective total amount of Synd4-positive cells. Horizontal lines indicate statistical significance (P < 0.05). C: quantification of PDGFR intensity per cell. Horizontal lines indicate statistical significance (P < 0.05). D: quantification of CD31 intensity per cell. Horizontal lines indicate statistical significance (P < 0.05). E: costaining of Synd4 in CD31-positive cells. After the total amount of CD31-positive cells was counted, as previously described, the respective Synd4-stained image was merged with the already merged CD31/Hoechst image. Cells with CD31/Synd4-positive staining were counted manually plus cells with Synd4 staining only. With the total amount of CD31-positive and Synd4-positive cells, the percentage of the costained cells was calculated. *P < 0.05.