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. 2021 Mar 1;6:10. doi: 10.1038/s41536-021-00120-8

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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. 2 — a, b Representative Verhoeff-Van Gieson stained sections and corresponding confocal fluorescence images of DAPI nuclei (blue) and Sca1-eGFP (green) expression in LCA after a 7 days and b 14 days. Scale bar = 20 µm. c The number of DAPI nuclei/hpf in cross sections of adventitia, media and intima following partial ligation of the LCA after 7 and 14 days compared to sham controls. Data are the mean ± SEM of 3–5 representative images per animal, n = 4. d The fraction of Sca1 cells/high power field (hpf) in the adventitia, media, and intima of sham and ligated vessels after 7 days and 14 days. Data are the mean ± SEM of 10–15 sections from four animals per experimental group. e Representative confocal fluorescence images of DAPI nuclei (blue), immunofluorescence of anti-eGFP (green) expression and anti-SMA (Red) in the LCA 14 days post-ligation in Sca1-eGFP mice. Data are representative of five images per animal. f Cumulative analysis of Sca1⁺ cells, SMA⁺ cells, double stained Sca1/SMA-positive cells and the percentage of Sca1⁺ cells that were SMA1⁺ in the adventitial, medial and intimal layer of RCA and LCA vessels after 21 days, respectively. Data are the mean ± SEM, n = 15–20 sections from 5 animals. g Representative confocal fluorescence images of DAPI nuclei (blue), immunofluorescence of anti-eGFP (green) expression and anti-Sca1 (far red white) in sham and ligated LCA after 21 days in S100β-eGFP mice. Scale bar = 20 µm. h Cumulative analysis of S100β⁺ cells, Sca1⁺ cells, double stained Sca1/S100β positive cells and the percentage of S100β⁺ cells that were Sca1⁺ in the adventitial, medial and intimal layer of RCA and LCA vessels, respectively after 21 days in S100β-eGFP mice. Data are the mean ± SEM, n = 15–20 sections from 5 animals/group. i. Representative confocal fluorescence images of DAPI nuclei (blue), immunofluorescence of anti-eGFP (green) expression and anti-SMA (far red, white) in the RCA and LCA 21 days post-ligation in S100β-eGFP mice. Data are from a minimum of four animals per experimental group. Scale bar = 20 µm. j Cumulative analysis of S100β⁺ cells, SMA⁺ cells, double stained S100β/SMA-positive cells and the percentage of S100β⁺ cells that were SMA⁺ in the adventitial, medial and intimal layer of RCA and LCA vessels, respectively, after 21 days in S100β-eGFP mice. Data are the mean ± SEM, n = 15–20 sections from 4 animals/group.