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. 2021 Sep 6;12:5270. doi: 10.1038/s41467-021-25653-w

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© The Author(s) 2021

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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Fig. 1 — a Violin plot showing the expression of Acta2 in CM1–CM5 cardiomyocyte populations. Expression levels are shown as reads per 10k counts. b Acta2 and Tnnt2 RNA transcripts detected by RNA-scope probes in the left ventricular myocardium on a transverse section of P1 hearts. Scale bar, 20 μm. c Quantification of Acta2 transcript levels in cardiomyocytes from P1 and P8 heart; n = 29–34 cardiomyocytes were quantified. d Acta2 and Tnnt2 RNA transcripts detected by RNA-scope probes in the left ventricular myocardium on a transverse section of P8 hearts. Scale bar, 20 μm. e Schematic diagram showing the spatial transcriptome analysis of heart sections to identify anatomic locations of cardiac cell type populations. f Hematoxylin and eosin (H&E) image of a heart transverse section collected at 7-day post P1 MI. Left and right ventricles are indicated. Black arrowheads depict fibrotic tissue; Scale bar, 500 μm. g Left: cell type composition shown in pie-charts for individual spatial spots mapped onto the heart section with H&E staining shown in (f). Right: the boxed region is shown in high magnification overlayed with the H&E image to highlight the fibrotic region; Scale bar, 200 μm. h–m Anatomic localization of endocardial cells (h), immune cells (i), fibroblasts (j), CM1 (k), CM5 (l), and CM4 (m) on the same heart section in (f). Color scale indicates cell type ratio from 0 to 1. Endocardium is outlined by black dash lines, and infarct region, which is identified by the location of suture, is outlined in white; Scale bar, 500 μm. n Expression correlation plots between Nrf1, Acta2, and Mki67 in individual cardiomyocytes from our previous snRNA-seq data (GSE130699). Nrf1 positively correlates with Acta2 (Spearman’s coefficient = 0.91). Color scale represents the expression level of Mki67. Expression levels are shown as reads per 10k counts. o, p Nrf1 and Tnnt2 RNA transcripts detected by RNA-scope probes in the left ventricular myocardium on transverse sections of P1 hearts (o) and P8 hearts (p). Scale bar, 20 μm. q Quantification of Nrf1 expression levels in cardiomyocytes from P1 and P8 hearts; n = 30 cardiomyocytes were quantified for each group. f–m experiments were repeated independently twice with similar results. ***p < 0.0001, Student’s t-test two-tailed. c and q arb. units. arbitrary units. e and g EC endothelial cells, EndoEC endocardial cells, FB fibroblasts, EPI epicardial cells.