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. 2020 Jul 17;10:11883. doi: 10.1038/s41598-020-68373-9

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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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Chronological expression of nodal markers in grafted human ES cell-derived cardiomyocytes (hESC-CMs) in injured hearts. hESC-CMs were transplanted on week after induction of myocardial infarction. Histological analysis was performed at the same time-points. (a–c) HCN4 (red) staining in the graft area. (d–f) SHOX2 (red) and human specific Lamin A+C (green) staining. SHOX2⁺/Lamin A+C⁺ cells (arrow heads) were designated as graft nodal cells. (g–i) Staining against β-MHC (red) and TBX3 (green). (j) Quantitative analysis of the number of SHOX2⁺/Lamin A+C⁺ cells divided by the number of total Lamin A+C⁺ graft cells (n = 5 per group). Data represent mean ± SEM. *P = 0.0036 vs. 2 weeks, + P = 0.0116 vs. 4 weeks by ANOVA with Tukey’s post hoc test. (k) Quantitative analysis of the number of TBX3 + cells divided by that of β-MHC⁺ graft cardiomyocytes (n = 5 per group). Data represent mean ± SEM. *P = 0.0003 vs. 2 weeks, + P = 0.0015 vs. 4 weeks by ANOVA with Tukey’s post hoc test.