Fig. 3

Increased sulfated monocytes/macrophages in CKD-related atherosclerosis. (A + D) Oil red O staining and quantification of lesions in the aorta en-face showed that atherosclerotic lesion size increased significantly in CKD-ApoE^−/− mice compared with Sham-ApoE^−/− mice. Scale bar: 2 mm. (B) Oil red O staining and Masson-trichrome (collagen in blue) staining of aortic sinus atherosclerotic lesions from CKD-ApoE^−/− and Sham-ApoE^−/− mice. The results are quantified in E + F, showing that CKD-ApoE^−/− mice have increased aortic sinus atherosclerotic lesions area but lower collagen area. Scale bar: 400 μm in the top image and 200 μm in the lower image. (C + G) Macrophages highlighted by immunohistochemical staining for CD68 increased significantly in aortic sinus plaques of CKD-ApoE^−/− mice compared with Sham-ApoE^−/− mice. Scale bar: 400 μm in the top image and 200 μm in the lower image. (H) Immunostaining showed co-localization of sulfotyrosine (green) and F4/80 (red) in aortic root sections, indicating the sulfated monocytes/macrophages in CKD-related atherosclerosis. Scale bar: 100 μm in the left image and 50 μm in the right image. (I + J) Peripheral blood was extracted from Sham-ApoE^−/− and CKD-ApoE^−/− mice and sulfated nucleated cells were determined by immunofluorescence staining with anti-sulfotyrosine antibody. The results showed that the percentage of sulfotyrosine-positive nucleated cells was significantly increased in CKD-ApoE^−/− mice. Scale bar: 50 μm. n = 5 for Sham-ApoE^−/−, n = 7 for CKD-ApoE^−/−. Data are expressed as means ± SD. ^**p < 0.01, ^***p < 0.001 compared with the Sham-ApoE^−/− group