Atherosclerosis Analysis in Murine Aortas

The aortic roots were isolated from mice, fixed with 4% paraformaldehyde, embedded in optimum cutting temperature compound (OCT), and cut into 5 µm-thick sections. Atherosclerotic lesions were detected by hematoxylin and eosin (HE) staining assay, and the lipid deposition was evaluated by Oil Red O staining kit (Nanjing Jiancheng Biology Engineering Institute, Nanjing, Jiangsu, China), as previously described^{39 (link)}. TUNEL staining and immunostaining of caspase-1 and CD31 (endothelial cell marker) on the aortic roots were performed to detect cell death and activation of caspase-1 in endothelial cells. The images were examined under a laser scanning confocal microscope (FV300, Olympus, Japan).