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. 2020 May 21;23(4):581–597. doi: 10.1007/s10456-020-09727-9

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — Distinct tumor EC and adjacent normal EC subpopulations in tumor-bearing liver tissues. a Characterization of HDD-induced intrahepatic tumors. (a) Gross image of liver harboring multiple HDD-induced tumors. Histological features of HDD-induced tumors by H&E (b), Masson Trichrome (c) and keratin 19 (Krt19) RNAScope (d) at × 20 magnification. b t-SNE plot of combined ECs collected from normal liver, dissected tumor, tumor-adjacent normal and undissected tumor-bearing liver (tumor and adjacent normal combined) tissue from C57BL/6 mice. Left, single cells colored by identified clusters; Right, cells colored by sample type. Bottom, nomenclature of sample types. c Heatmap of top 15 EC cluster-specific genes. Clusters (corresponding to the t-SNE plot in 2b) were annotated based on both known EC marker genes and sample type. d RNAScope on intrahepatic tumors and adjacent normal liver tissues of SCID mice showing tumor-specific expression of Aplnr and upregulation of Selp in sinusoid and PV (see naïve normal tissue expression in Fig. 1f for comparison). Pecam1 was used as pan-EC marker. e. Gene and UMI counts across clusters (corresponding to t-SNE plot in 2b). f Functional enrichment (GO terms) of genes preferentially expressed (upregulated) in tumor ECs compared to non-tumor liver ECs. g Feature plot showing upregulation of Lrg1 and Sema3d in adjacent normal ECs compared to naïve ECs (corresponding to EC cluster t-SNE plot in b)