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. 2024 Feb 7;626(8000):864–873. doi: 10.1038/s41586-023-06950-4

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© The Author(s) 2024

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 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. 4 — a,b, RNA-seq analysis of KCs from Clec4f^creId3^f/f mice (n = 2) and Id3^f/f littermates (n = 3). a, The pathways downregulated in Clec4f^creId3^f/f mice. Pos., positive. b, Selected differentially expressed genes. c, RT–qPCR analysis of selected genes in KCs from Clec4f^creId3^f/f mice and Id3^f/f littermates. n = 3 per group. d, Flow cytometry analysis of the expression of SIRPA and dectin-1 by KCs from Clec4f^creId3^f/f mice and Id3^f/f littermates 2 weeks after intraportal injection or not of 1 × 10⁶ KPC-1-GFP cells. n = 3 per group. Ctrl, control. e, The percentage of TIM4⁺ KCs containing GFP in the livers from Clec4f^creId3^f/f mice (n = 4) and Id3^f/f littermates (n = 3) treated as described in d. f, Immunofluorescence analysis of the percentage of TIM4⁺ KCs stained with GFP in Clec4f^creId3^f/f mice and Id3^f/f littermates 24 h after intraportal injection of 1 × 10⁶ KPC-1-GFP cells (left). n = 4 per group. Right, the number of CD45⁻GFP⁺ tumour cells per liver lobe was analysed using flow cytometry in the same mice. n = 5 per group. g, Engulfment of KPC-1-tdT cells by Clec4f^creId3^f/f or Id3^f/f KCs in vitro, as in Fig. 2g. The plots show the percentage of KCs engulfing Cas-Green⁻ KPC-1-tdT cells (n = 3 per group), and the time from stable interaction between KCs (n = 25 (Id3^f/f) and n = 7 (Clec4f^creId3^f/f)) and tumour cells to engulfment or caspase-3/7 cleavage. h, Expression of chemokines and cytokines by TIM4⁺ KCs from Id3^f/f and Clec4f^creId3^f/f mice treated as in d. n values are indicated. i,j, Flow cytometry (i) and immunofluorescence (j) analysis of CD8⁺ T cells and LAMP1⁺NKP46⁺ cell numbers per g of liver or per mm² in mice from h. n = 5 mice per group. k, IFNγ and TNF expression by NKP46⁺ and CD8⁺ T cells from i and j. l, The liver tumour burden was analysed using photoradiance 2 weeks after injection of 1 × 10⁶ KPC-1-luci cells in Id3^f/f mice treated with IgG (n = 6) or anti-CD8/NK1.1 (n = 3) and Clec4f^creId3^f/f mice treated with IgG or anti-CD8/NK1.1. n = 4 per group. Statistical analysis was performed using unpaired two-tailed t-tests (c, d, f, g, i, k and l), one-way ANOVA (e, j and l) and Kruskal–Wallis tests (h). Data are mean ± s.d. Norm., normalized.

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