Fig. 1

High fat diet (HFD) reduces vascular and plasma 5-MTP via TLR2. A Paraffin-embedded aortic tissues were prepared from chow diet–fed ApoE^−/− (n = 6), HFD-fed ApoE^−/− (n = 8), and ApoE^−/−Tlr2^−/− (n = 9) mice for 20 wk and the 5-MTP contents were analyzed by immunohistochemistry as shown in the upper two panels. Insets in the second panel show the magnified views taken from the areas of aortic tissues in the first panel. Vascular chondrogenesis and calcifications in the bottom two panels were visualized with Alcian blue and ARS (ARS) staining, respectively. 5-MTP immunopositive areas in (B) the total artery, C aortic medial layer, and D intimal layer were measured using ImageJ software and expressed as the percent positive area out of the total tissue area. E Plasma 5-MTP levels in mice fed a chow diet (n = 9) or HFD (n = 20) for 20 wk were measured by quantitative LC–MS-MS. F, G TPH-1 contents in paraffin-embedded aorta tissues from mice fed a chow diet (n = 5) or HFD (n = 7) for 20 wk were analyzed by immunohistochemistry. Negative control represents staining with a 2nd antibody. Representative microphotographs are shown in (F) and (G) Immunopositive areas in the aorta tissues were quantified using ImageJ software as a percentage of the total aortic area in each section. Data in B–E and G represent mean ± SEM. Lu lumen, Neo neo-intima, Med media, An adventitia. P < 0.05, P < 0.01, P < 0.001 determined by t-test or a one-way ANOVA