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. 2020 Oct 20;11:5300. doi: 10.1038/s41467-020-19150-9

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© The Author(s) 2020

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. 6 — a and b Genetic tracing of Cx40+ cells labeled at E18.5 by Tam injection. Confocal images of whole-mount CNTN2-immunostaining of Cx40-CreERT2::R26R-YFP opened-left ventricle at P21. a″ and b″, Immunostaining for YFP and CNTN2 on control, or Nkx2-5^+/− mice shows E18.5 Cx40+-derived cells that participate to the PF network. Scale bars = 1 mm; a’ and b’ are high magnifications of left bundle branch; a″ and b″ are high magnifications of PF. c–e Strategy to induce conditional deletion of one copy of Nkx2-5 allele (Nkx2-5^flox/+) using Cx40-CreERT2 line as inducible Cre driver. Deletion is performed at E18.5 when Cx40+ cells are mostly PF cells or E10.5 before the massive recruitment of late progenitors. R26R-YFP reporter line is used to compare the genetic tracing of Cx40+-derived cells within P21 control, Nkx2-5^+/− and Nkx2-5^flox/+ hearts (N = 7 of each genotype for E18.5 and N = 3 for E10.5). e Confocal imaging of left PF network. Scale bars = 500 µm; arrowheads indicate non-conductive Cx40+-derived cells.