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Fig. 2 | Journal of Translational Medicine

Fig. 2

From: Bilirubin ameliorates murine atherosclerosis through inhibiting cholesterol synthesis and reshaping the immune system

Fig. 2

Bilirubin inhibits atherosclerotic lesions formation and hepatic lipid accumulation in ApoE−/− mice. ApoE−/− mice fed with a western-type diet were intraperitoneally injected with (bilirubin, n = 14) or without (control, n = 8) bilirubin. After ten weeks, the animals were sacrificed and the aorta, heart, brachiocephalic artery, and liver tissue were collected for morphometry. A Aortas fixed with formalin was performed en face Oil Red O (ORO) staining and the lesion area was quantified. B Series of frozen sections of aortic sinus were subjected to hematoxylin–eosin (H&E), ORO, and Masson staining. Aortic sinus lesion area was quantified in the context of H&E staining. C Histological analysis of brachiocephalic arteries after H&E and ORO staining. The neointima/media ratio was calculated. D Immunofluorescence staining was applied to analyze the distribution of Galectin-3 ( +) (macrophages, red) and α-SMA ( +) (smooth muscle cells, green) in aortic valve lesions. The area of Galectin-3 ( +) and α-SMA ( +) was quantified. E Histological analysis of liver after H&E and ORO staining. Area with oil red color are steatosis area. The black arrows indicate the representative steatosis area. The red arrow indicates the normal area. F The amount of total cholesterol and free cholesterol in liver lysates was measured. Cholesteryl esters were calculated by subtracting the free cholesterol values from the total cholesterol value. Error bars represent mean ± SD. n (control) = 8, n (bilirubin) = 14. *P < 0.05, **P < 0.01 and ***P < 0.001. Differences were determined by an unpaired t-test

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