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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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Increased recruitment of Mpx-positive neutrophils at the onset of regeneration after multiple cryoinjuries. (A) Cross sections of adult zebrafish hearts immunostained for L-plastin (green) and Myeloperoxydase (Mpx, red) at 4 dpci following one, two three and six cryoinjuries (CIs). The cardiac muscle is detected by F-actin staining (Phalloidin, blue). Both L-plastin and Mpx were detected in the wound. L-plastin/Mpx double positive cells are indicated (white arrow). The dashed line encircles the post-injury zone. Scale bars = 100 µm. (B) Analysis of the percentage of L-plastin-positive area within the wound. No significant change (NS) is observed between the experimental groups. N ≥ 3. Scatter plot of the data with a large bar indicating the mean and smaller bars representing the SEM. (C) Analysis of the percentage of Mpx-positive area in the wound. Multiple cryoinjuries significantly increase the number of neutrophils compared to specimens after one cryoinjury. N ≥ 3; *P < 0.05. Scatter plot of the data with a large bar indicating the mean and smaller bars representing the SEM.