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. 2023 Sep 9;14:5567. doi: 10.1038/s41467-023-41279-6

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Force-directed layouts of 10,354 endocardial and endocardial-derived cells displaying timepoint, annotated cluster, and origin. b A dot plot displaying select genes enriched within endothelial-derived clusters, including VEC populations and AV mesenchyme. Size of the dot represents proportion of the cluster that expresses each gene. Colour indicates level of expression. c Force-directed layouts of 10,354 endocardial and endocardial-derived cells displaying pseudotime and differentiation (diff.) potential. Arrows indicate high diff. potential in domains of endothelial-mesenchymal plasticity (EMP). d Gene trends along pseudotime show bifurcation between VEC and AV mesenchymal trajectories during the endocardial-to-mesenchymal transition. e Differentiation potential as a function of pseudotime for endothelial-derived lineages progressing toward a VEC terminus. Each dot represents a single-cell that is colour-coded by embryonic timepoint or cluster. f Immunostaining of AV canals for SOX9, ERG, and NFATc1 shows co-expression of on the flow side of the valves (white arrowheads) at E10.5 (n = 10) and E11.5 (n = 4). Scale bars: 100 µm, 25 µm in regions of interest (ROIs). AV atrioventricular, endo. endothelium, mes. mesenchyme. See also Supplementary Fig. 2 and Supplementary Data 2.