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. 2020 Jul 9;10:11404. doi: 10.1038/s41598-020-68223-8

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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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Cardiac PW1⁺CD51⁺ cells express a fibrogenic program. (A) Hierarchical clustering based on the expression of Peg3, Itgav, and known cell type markers in FACS-isolated PW1⁺ cells, cardiomyocytes, and non-myocyte fractions from normal mouse hearts. (B) Hierarchical clustering showing high expression of collagen genes in PW1⁺ cells. (C) Expression of Peg3 (yellow) and Itgav (blue) in major cardiac cell populations identified after unsupervised clustering of single-cell RNA-sequencing data as previously reported²⁵. Each point depicts a single cell. Principal component analysis and t-distributed stochastic neighbor embedding (t-SNE) were performed to provide a 2D mapping of clusters. Joint expression of Peg3 and Itgav within single cells is showed in purple. (D) The abundance of Peg3 and Itgav mRNAs and those encoding Itgβ1, Itgβ3, Itgβ5, and Itgβ8 according to cell types identified using single-cell RNA-sequencing from normal mouse hearts. (E) Western blots for ITGβ1, ITGβ3, ITGβ5, and ITGβ8 in PW1⁺ versus PW1⁻ cardiac fractions. Proteins from HUVEC cells and neonatal mouse heart were used as positive controls. Ponceau Red staining showed equal protein loading.