Fig. 1: scRNA-seq analysis of Twist1-regulated EC clusters.

Male Twist1^ECKO (Twist1^fl/fl Cdh5^CreERT2/+ ApoE^−/−; n = 4) and control (Twist1^fl/fl Cdh5^+/+ ApoE^-/--; n = 5) mice aged 8 weeks were fed a Western diet for 8 weeks to induce atherosclerotic lesions. Tamoxifen was then administered for 5 consecutive days to induce Twist1 deletion in experimental mice, and a Western diet was provided for an additional 6 weeks. Aortas were analysed by FACS of pooled CD31⁺ CD45⁻ cells coupled to scRNA-seq. A t-SNE map of single-cell RNA sequencing from Twist1^ECKO and control mice, colored by cluster assignment. Clusters were identified using unbiased hierarchical clustering. B t-SNE map showing the cell contribution of Twist1^ECKO and control mice to each subpopulation. C Bar graphs showing the absolute number of cells from Twist1^ECKO and control mice in each cluster (left), and the proportion of Twist1^ECKO and control cells in each cluster (right). The dotted line indicates the 50% threshold. Clusters 2, 7 and 8 are largely composed of ECs derived from control mice, whereas clusters 4 and 5 are mainly composed of ECs derived from Twist1^ECKO mice. D Average levels of EndMT markers and EC markers (AUCell score) were calculated for individual cells and are presented as violin plots. The white dot represents the median. The box is the interquartile range (IQR) (the upper limit of the box is quartile 3 (Q3), the lower limit is quartile 1 (Q1)). The thin lines (whiskers) extend to the lower and upper adjacent values (Q1 −  1.5 IQR and Q3 + 1.5 IQR). Clusters 2, 3, 7 and 8, which are highlighted with a blue box, show enriched expression of EndMT markers.