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. 2021 Jul 6;12:4155. doi: 10.1038/s41467-021-24414-z

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© The Author(s) 2021

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. 3 — a FISH using probes against Gfp (green) to detect epicardium-derived cells and Sema3d (white) or Slit2 (red) reveals diverging localization of vascular guidance cues within the epicardium and interstitium between embryonic (E)12.5 and E16.5. a′, 3× zoom of E16.5. b Expression of Sema3d (red) in Wt1 lineage-derived cells (Gfp⁺, green) relative to Pecam1⁺ (platelet endothelial cell adhesion molecule-1, white) endothelial cells. b′, 3× zoom of E16.5. c Expression of Slit2 (red) in Wt1 lineage-derived cells (Gfp⁺, green) relative to Pecam1⁺ (white) endothelial cells. c′ 3× zoom of E16.5. Yellow arrowhead, Gfp⁺/Sema3d⁺ cells. Orange arrowhead, Gfp⁺/Slit2⁺ cells. Dashed yellow line represents the endocardium–myocardium border. DAPI staining was utilized to visualize nuclei (blue). Scale bar, 20 μm. Scale bar 3× zoom, 10 μm. Endo endocardium.