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. 2021 May 25;7:36. doi: 10.1038/s41421-021-00271-4

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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. 3 — a UMAP plot of acinar cells and ductal cells in dense- and loose-type PDACs. b Proportion of cells from acinar and ductal cells in dense- and loose-type PDACs. c Violin plots of selected marker genes from clusters 0, 2, and 6, showing normalized expression in the different clusters. d KEGG and GO analysis comparing cluster 6 against cluster 2. e Reclustering of T-cell types in the data set (clusters 4, 5, and 19 from Fig. 1c), represented as a UMAP plot. f Proportion of T-cell subpopulations in dense- and loose-type PDACs (four dense cases, three loose cases). The proportion of the CD8⁺ T-cell group in all T cells is significantly increased in loose-type PDAC. The y axis represents the proportion of each T-cell subgroup in all T cells, and the x axis represents different T-cell subgroups. Red column and green column represent dense-type PDAC and loose-type PDAC, respectively. Data are presented as means ± SEM. All statistical analyses were performed with the two-sided Mann–Whitney U test. *P < 0.05. g GSEA showing enriched pathways in CD8⁺ T cells comparing loose-type against dense-type PDAC. h Multiplex immunofluorescence staining was conducted in nine PDAC samples undergoing scRNA-seq to confirm the correlation between CAF subpopulations and infiltrating CD8⁺ T cells (n = 9, POSTN for myCAF, PLA2G2A for meCAF, CD8 for CD8⁺ T cell). Representative immunofluorescence results showed much more infiltrating CD8⁺ T cells in loose-type PDAC than in dense-type PDAC. i Quantification of CD8⁺ T cells for n = 7 cases (four dense cases and three loose cases), including representative case. The y axis represents percentage of CD8⁺ T cell per visual field (40×), and the x axis indicates dense-type PDAC and loose-type PDAC, respectively. Data are presented as means ± SEM. All statistical analyses were performed with the two-sided Mann–Whitney U test. *P < 0.05. j GSEA showing enriched pathways in macrophages comparing loose-type against dense-type PDAC.