JCAD deficiency reduces atherosclerosis in ApoE−/− mice fed a western-type diet. (A-B) En face Oil Red O–staining of whole aorta from male (A, n = 15–16) and female (B, n = 7–12) JCAD+/+; ApoE−/− and JCAD−/−; ApoE−/− mice after 12 weeks of western-type diet feeding. Quantification of en face lesion area. Data were represented as percent Oil Red O positive area of the entire aorta measured by Image J. Images presented were from a composite of 2–3 images from the same aorta. (C) Representative Oil Red O–stained aortic sinus cryosections from male JCAD+/+; ApoE−/− (n = 6) and JCAD−/−; ApoE−/− (n = 5) mice. Quantification of aortic sinus lesion area between two groups. Data were represented as percent Oil Red O positive area of the entire sinus measured by Image J. (D–F), Analysis of plaque composition in aortic sinus. The contents of macrophages (CD68 positive, green, D), smooth muscle cells (α-SMA positive, red, E) were analyzed by immunostaining-based quantification of the positive signal (mean fluorescence intensity in arbitrary units) was provided in right panels, n = 6 per each group, scale bar = 100 μm. Collagen content (blue, F) was determined by Masson’s trichrome staining. Quantification data were presented as % lesion area in the right panel, n = 8 for JCAD+/+; ApoE−/− and n = 7 for JCAD−/−; ApoE−/− group. α-SMA, alpha-smooth muscle actin.