Figure 5.

Atheroprotective effects of Ccn2 are exerted through SMC-dependent mechanisms. (A) Experiment design (n = 9/7/9/8 for chow-fed Ccn2^wt/wt and Ccn2^SMCΔ/Δ mice and wd-fed Ccn2^wt/wt and Ccn2^SMCΔ/Δ mice, respectively, after excluding five mice that met the pre-specified humane endpoints (see supplementary material online, Figure S7D). In addition, four Ccn2^SMCΔ/Δ mice were sacrificed after 12 weeks of wd feeding to enable phenotype assessment at this time point. (B, C) Mouse weight (B) and plasma cholesterol (C) through the course of the experiment. (D, E) Ex vivo images of Ccn2^wt/wt and Ccn2^SMCΔ/Δ aortas. (F) Mass of aortas relative to body weight. (G) Aorta length (beginning of aortic arch to iliac bifurcation). (H) Oil Red O staining of en face-prepared thoracic aortas. Scale bars = 2.5 mm. (I) Percentage of en face-prepared thoracic aorta area stained positive for lipid. (J) Area of en face-prepared descending thoracic aortas. (K, L) Magnification of lipid-stained areas of descending thoracic aortas from Ccn2^wt/wt mice after 24 weeks of hyperlipidaemia (K) and Ccn2^SMCΔ/Δ mice after 12 weeks of hyperlipidaemia (L). Arrowheads in point to ostia of aortic branches. (M + N) Histological images of Oil Red O-stained lesions (top) and the same specimens stained for Lgals3 (red), Acta2 (green), and nuclei. m = media. Scale bars = 50 µm. Data in B and C were analysed by two-way ANOVA. Data in F and G and I and G were analysed by unpaired t-test with Welch’s correction or Mann–Whitney U test.