Fig. 3: Lineage tracing of the SHF and NC aortic arch highlights respective contributions to the arch regional signature.

a, Experimental design. Single-cell suspensions were generated from isolated aortic arches of mice expressing eGFP after Mef2c-Cre recombination (SHF-derived cells) or tdTomato after recombination with Wnt1-Cre (NC derived). Lineage-traced cells were bioinformatically segregated from the data using transcript expression of the indicated fluorescent proteins before downstream analysis. b, Representative IF images of NC-vSMCs (green) after Wnt1-Cre-mediated recombination visualized in the adult mouse aorta, n = 6. c–f, Violin plots for expression of eGFP (c), tdTomato (d) and vSMC contractile markers (Myh11 (e), Mylk (f)) in the Mef2c versus Wnt1 datasets. g, UMAP plot visualizing the distribution of Mef2c⁺ and Wnt1⁺ lineage-tracing experiments and their respective controls. Ref, reference. h, Heatmap of the top 45 genes differentially expressed between eGFP⁺ Mef2c-derived vSMCs and tdTomato⁺ Wnt1-derived vSMCs. Gm42418, Rn18s-rs5 (gene symbol); Olfr1033, Or5m3b (gene symbol). i, Gene ontology enrichment for arch-derived vSMCs highlighting signature genes from each of the two developmental origins calculated via hypergeometric distribution. P values for enrichment categories are as follows: vascular development, 3.88 × 10⁻⁹; NABA core matrisome, 5.53 × 10⁻⁹; heart development, 2.63 × 10⁻⁸. j–o, Violin plots visualizing the expression of lineage-derived vSMC markers (Clic4 (j), Tnnt2 (k), Tbx20 (l), Cacna1d (m), Etv1 (n), Fgf2 (o)) in aortic arch Mef2c⁺ and Wnt1⁺ vSMCs. Panel a created using BioRender.com.