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. 2021 Jan 15;11:1520. doi: 10.1038/s41598-020-80848-3

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Gata4, Mef2c, and Tbx5 expression in the beating and non-beating groups. (a) Left three panels, gene expression levels in the beating and non-beating group. Right panel, gene expression levels in the beating and non-beating group relative to that of β-actin. (b) Representative immunofluorescence images of a cluster of round cells expressing Gata4 and Mef2c on day 14. (c) Phase-contrast and immunofluorescence images of beating cells expressing cardiac troponin T (cTnT) and Mef2c on day 28. The arrowhead indicates a beating cell. Scale bar, 100 µm. DAPI, 4′,6-diamidino-2-phenylindole, ** p < 0.01; **** p < 0.0001 by t-test or ANOVA and Tukey’s post hoc test. (d) The frequency of round cells expressing Gata4 and Mef2c (10 round cells were examined per experiment, n = 13). (e) The frequency of beating cells expressing Mef2c and cTnT (15 beating cells were analysed per experiment, n = 4).