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. 2020 Feb 20;10:3095. doi: 10.1038/s41598-020-59868-6

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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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Cox2a expression is highest in epicardial cells at 3 dpa. (A–C) Representative FACS dot plots displaying gating parameters used to isolate cells from (A) Tg(cmlc2:EGFP); Tg(tcf21:DsRed), (B) Tg(fli1a:eGFP), (C) Tg(mpeg1:YFP) reporter lines at 3 dpa. (D) qPCR analyses of population specific markers within reporter(+) and reporter(−) cells validate the purity of FACS isolated cells. (mean ± s.e.m. n = 3 biological replicate for each cell type. Student’s t-test *P < 0.05) (E) qPCR studies in 3 dpa FACS sorted cells showed cox2a expression is significantly higher in tcf21(+) cells, relative to all other cell types examined. (F) qPCR of FACS sorted cells showed that at 3 dpa, cox2b expression was significantly higher in fli1a(+) cells relative to all other cell types examined. (G) There was no significant difference in cox1 expression among the resident cardiac cells assayed. cox1 expression in mpeg1(+) cells was significantly lower than that observed in fli1a(+) and tcf21(+) cells. Gene expression was calculated relative to cmlc2(+) cells. (mean = ±s.e.m. n = 2–5 biological replicates; 12–45 pooled ventricles per replicate. One-way ANOVA followed by Tukey’s multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001).