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. 2022 Oct 5;8:101. doi: 10.1038/s41421-022-00445-8

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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. 3 — a t-SNE plot showing mEPCs colored by clusters (n = 13). b Pie charts showing the relative abundance of mEPC clusters (colored in line with a) across tumor samples (n = 16, excluding two samples with EPC counts < 30). c Heatmap showing three subtypes of mEPCs classified by hierarchical clustering based on GSVA analysis (MSigDb_C6 oncogenic signatures), with corresponding clinicopathological features displayed. d Heatmap showing top-ranking DEGs for three subtypes of mEPCs. e Heatmap showing the assignment of scRNA-seq-defined mEPC subtypes by analyzing the bulk RNA-seq data from an external GBC cohort (n = 111), based on GSVA analysis (MSigDb_C6 oncogenic signatures). P values for pathways with a significant enrichment are displayed (one-way ANOVA test; **P < 0.01, ***P < 0.001). f–h Representative IHC staining images of markers for subtype I (MUC2, OLMF4), subtype II (CTSD, BST2), and subtype III (MSLN, SPP1) by using our GBC tissue microarray cohort (n = 49), respectively. Scale bars, 500 μm, 5×; 200 μm, 20×. i–k Kaplan–Meier curves of overall survival when stratifying patients by high or low expression of mEPC subtype markers. P values were calculated by log-rank test.