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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. 4 — a Scheme illustrating the genetic tracing strategy using R26R-Confetti multicolor reporter mouse line crossed with non-inducible Mesp1-Cre mice. b Recombination of the confetti allele occurs during the short time-window of Mesp1 expression, between E6.25 and E7.75. Mosaic tracing analysis in control and Nkx2-5^+/− mice is performed at P21. c Whole-mount fluorescence views of Mesp1-Cre::R26R-Confetti opened-left ventricles at P21. Immunostaining for Contactin-2 (CNTN2) is used to label the mature PF network. Small panels (1–4) show high magnification confocal images of different PF network regions. Scale bars = 1 mm and 100 µm in insets. d Small panels (1–4) show high magnification confocal images of the PF network. Panels (1′–4′) are image reconstructions in which conductive only confetti+ cells are manually drawn. Non-conductive confetti+ cells are not represented. In control hearts, conductive confetti+ cells form either small or large monochromatic clusters. Large clusters (arrows) contribute to complex parts of the PF network. In Nkx2-5^+/− hearts, only small clusters contribute to the hypoplastic network. Scale bars = 1 mm and 200 µm in insets. e and f Dot plots of individual Mesp1+ confetti colors showing percentage contribution to the working myocardium versus PF network in control (WT) or Nkx2-5^+/− (NK) hearts.