Quantification of aortic atherosclerotic lesions

Aortas and aortic valves were prepared as follows. In brief, en face preparations of entire aortas were dissected, fixed, opened longitudinally and pinned on black wax plates. Then, lesions were visualized by staining with Oil Red O (Sigma). The aortic roots were fixed in 4% formaldehyde, processed and embedded in optimum cutting temperature compound. The resultant aortic sinus cryosections (7 μm) were stained with Oil Red O and haematoxylin. The mean atherosclerotic areas were calculated from eight different mice. Ten serial cryosections/tissue sections for each mouse were evaluated. The total atherosclerotic area for each plaque area measurement from each mouse was used for this calculation. image‐pro plus 6.703 software (Media Cybernetics) was used for statistical analysis.