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. 2020 Jul 14;10:11551. doi: 10.1038/s41598-020-68200-1

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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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Comparison of regenerative dynamics between hearts after multiple cryoinjuries. (A) Representative sections of regenerating hearts stained with the AFOG reagent, showing the myocardium (beige), fibrin (red) and collagen (blue) at different time points following cryoinjuries. At 4 and 7 dpci, the wound contains markedly more collagen after multiple cryoinjuries as compared to that after one cryoinjury. Scale bar = 100 µm. (B) Linear representation of the percentage of wounded area per ventricle sections at different regenerative time-points. All experimental groups exhibit similar dynamics of regeneration, but hearts after six cryoinjuries comprise the largest wound as compared to other groups, at 60 dpci. N ≥ 4 hearts, 3 sections per heart. (C) Histogram showing the percentage of collagen staining within the remaining wounded area. After multiple cryoinjuries, the amount of collagen is high already at the early time points of regeneration. N ≥ 4 hearts, 3 sections per heart.