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. 2023 Mar 31;14:1796. doi: 10.1038/s41467-023-37409-9

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© The Author(s) 2023

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. 5 — a Representative images and quantification showing the significantly decreased migration of endothelial cells (labeled with RFP) in E13.5 Nfatc1-Cre;Ptbp1^fl/fl;Rosa26-tdTomato mice compared to Nfatc1-Cre;Rosa26-tdTomato mice, which were assessed by the percentage of endothelial cell coverage on the ventral side of the heart. Scale bars, 200 µm. n = 5 mice for Nfatc1-Cre;Rosa26-tdTomato group, n = 6 mice for Nfatc1-Cre;Ptbp1^fl/fl;Rosa26-tdTomato group. b Immunofluorescence staining of EMCN in heart sections from E11.5 Ptbp1^fl/fl and Nfatc1-Cre;Ptbp1^fl/fl mice. Scale bars, 100 µm. The graph shows the thickness of the compact myocardium (CM) and trabecular myocardium (TM) of E11.5 Ptbp1^fl/fl and Nfatc1-Cre;Ptbp1^fl/fl mice. n = 3 mice per group. c Schematic diagram showing the experimental strategy of explant culture. d Immunostaining for cTnT (red) and ERG (cyan) in ventricular explants from Ptbp1^fl/fl and Nfatc1-Cre;Ptbp1^fl/fl mice after 2 days culture. Scale bars, 200 µm. The arrowheads indicate decelerated endothelial cell migration in the explanted tissues. e Left: immunostaining for NKX2.5 (red) and EdU (green) in Ptbp1^fl/fl and Nfatc1-Cre;Ptbp1^fl/fl ventricular explants cultured for 2 days. Scale bars, 25 µm. Right: Quantification of the EdU-positive cardiomyocytes in ventricular explants. n = 4 explants per group. f Representative images and quantification of ventricular explants showing the migration of endothelial cells (labeled with ERG) at 5 days after culture. The arrows indicate the direction of endothelial cell sprouting. Scale bars, 200 µm. n = 4 explants per group. g Representative images and quantification of the in vitro scratch assays in primary human umbilical endothelial cells (HUVECs) after PTBP1 knockdown (LV-sh-PTBP1). Scale bars, 200 µm. n = 3 independent experiments. BF bright field, TM trabecular myocardium, CM compact myocardium, LV left ventricle, RV right ventricle, Ctrl control lentivirus-shRNA, LV-sh-PTBP1 lentivirus-shRNA-PTBP1. The data are presented as the mean ± s.e.m. P values were calculated by unpaired two-tailed t test. Source data are provided as a Source Data file.