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. 2022 Sep 20;13:5511. doi: 10.1038/s41467-022-33052-y

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© The Author(s) 2022

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 Study design. EC endothelial cell, HR hormone receptor, HER2 human epidermal growth factor receptor 2, VD Viability Dye, pME peri-tumoral microenvironment, pEC peri-tumoral endothelial cells, TME tumor microenvironment, TEC tumor endothelial cells. The numbers related to FACS-sorting indicate sequence of sorting, color coded to indicate that each patient yields 4 samples (pME, pEC, TME, and TEC). b Patient characteristics. First line, patient identifier; second line, pathological tumor (pT) and nodal (pN) stage; third line, IUCC (International Union for Cancer Control) cancer stage according to the 8th edition; fourth line, hormone receptor status, coded by color for receptor type (gray, estrogen receptor; green, progesterone receptor) and bar length for Allred score (routinely used immunohistochemistry score based on the percentage of positive cells and the intensity of that staining); fifth line, differentiation grades; grade 1, well differentiated; grade 2, moderately differentiated (patient: #2–8); grade 3, poorly differentiated (patient: #1); sixth line, Ki67 proliferation index based on the clinically performed immunohistochemistry stainings. c UMAP-plot, showing the subclustering of 8433 endothelial cells (ECs) from tumoral (n = 8) and matched peri-tumoral (n = 7) breast cancer (BC) patients. LS lower sequencing depth, PCV post-capillary venules. d Heatmap of the expression levels of the top-10 marker genes in all EC subclusters. Color scale: red – high expression, blue – low expression. e Hierarchical clustering analysis of EC subclusters. Color differences in the dendrogram indicate clusters that were resolved by multiscale bootstrapping, p-value cutoff 0.4. Approximately unbiased (AU) p-values, and bootstrap probability (BP) values are indicated for each dendrogram branch in purple and green, respectively. f UMAP-plots of all 8433 ECs color-coded by condition. Dotted lines surround angiogenic (left) and capillary (right) EC subclusters. g Abundances of EC subclusters across conditions (pEC (gray), TEC (red)). Left panel: y-axis depicts % of total enriched ECs, x-axis depicts EC subtypes color coded as in panel (c). Right panels: similar representation of the data as in left panel, but EC subtypes were pooled as indicated. Data are mean ± SEM, n = 7 for pEC, n = 8 for TEC, *p < 0.05, **p < 0.01 (exact p-values = 0.008, 0.0063, 0.0498, respectively (left panel), and 0.0151, 0.0021, respectively (right panel)), paired t-test (two-tailed) per subcluster (taking only into account the 7 complete pairs).