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Figure 2 Sclerotic changes of aortic valves of senile wild-type and apolipoprotein E (ApoE)–/– mice. (A) Gross appearance of aortic valves of 96-week-old wild-type mice and 95-week-old ApoE–/– mice. Scale bar, 500 µm. (B) Hearts were taken from 94- to 98-week-old wild-type (male n = 2, femalen = 1) and 74- to 97-week-old ApoE–/– (male n = 5, female n = 5) mice, and embedded in OCT compound. Frozen sections containing aortic valves were stained by von Kossa method (VK) to detect ectopic calcification. Antiosteocalcin (OCL) immunostaining was performed on the frozen sections obtained from wild-type and ApoE–/– mice. Arrows indicate the positive area. Scale bars, 100 µm (upper panel) or 20 µm (middle and lower panels). (C) Immunohistochemical studies were performed on the frozen sections. Endothelial cells were detected by anti-CD31 immunostaining (CD31). Arrowheads indicate the sites of endothelial denudation. Macrophages and T cells were detected by immunostaining with anti-MOMA-2 and anti-CD3 antibodies, respectively. Smooth muscle-like cells were identified using an anti- -smooth muscle actin (SMA) antibody. Arrows indicate the positive area. Scale bar, 20µm.
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