Fig. 7: Twist1 promotes aortic root plaque growth and features of stability in vivo.

Male Twist1^ECKO (Twist1^fl/fl Cdh5^CreERT2/+ ApoE^−/−) and control mice (Twist1^fl/fl Cdh5^+/+ ApoE^−/−) aged 8 weeks were fed a Western diet for 8 weeks to induce atherosclerotic lesions. Tamoxifen was then administered to both groups for 5 consecutive days to induce Twist1 deletion in ECs of experimental mice. Mice were then fed a Western diet for an additional 6 weeks (totaling 14 weeks of Western diet). Paraffin-embedded sections of aortic roots were stained with A Hematoxylin and Eosin (H&E) to quantify the plaque size in Twist1^ECKO (n = 10) and control (n = 9) mice, B Picrosirius Red (visualized under polarized light) to quantify collagen content in Twist1^ECKO (n = 10) and control (n = 10) mice, C antibodies against ACTA2 to quantify vSMCs content in Twist1^ECKO (n = 10) and control (n = 10) mice, D antibodies against MAC3 to quantify macrophage content in Twist1^ECKO (n = 10) and control (n = 10) mice, E Hematoxylin and Eosin (H&E) to quantify necrotic core (highlighted in yellow) content in Twist1^ECKO (n = 10) and control (n = 9) mice. Representative images are shown (Scale bar = 100 μm). Mean values are shown ± standard errors. Blue dashed line represents a baseline value measured in a cohort of Twist1^+/+ Cdh5^+/+ ApoE^−/− mice exposed to a Western diet for 8 weeks (see Fig. S21). Differences between means were analysed using a one-sided unpaired t-test. * P < 0.05; ** P < 0.01; *** P < 0.001. Source data are provided as a Source Data file.