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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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Extended Data Fig. 2 — a-g- Representative immunofluorescence staining for Clec4f, F4/80, Tim4, tdT, CK19 on frozen liver sections from wt C57BL/6j mice 8 weeks after pancreatic orthotopic injection of 2 × 10⁵ KPC-2-tdT cells. b-g- idem, from wt C57BL/6j mice 2 weeks after intra-portal injection of 1 × 10⁶ KPC-1-tdT cells, 1 × 10⁶ KPC-1 cells, 5 × 10⁵ B16F10-luci-tdT cells, 1 × 10⁶ LLC1-luci-tdT cells, 1 × 10⁶ Pan02-luci-tdT cells, or 1 × 10⁶ MC38 cells. n = 3 mice per group. h- Flow cytometry analysis of the % of CD45.1⁺ cells among Tim4⁺CD206⁺ KCs and Tim4⁺CD206^hi KCs in the liver of CD45.1/CD45.2 parabiotic pairs 2 weeks after intra-portal injection of 10⁶ KPC-1-tdT in the CD45.2 partner (n = 5), or not injected as control (n = 4). i- table represents the percentage (mean and sd) of CD45.1⁺ partner-derived cells among Tim4⁺, Tim4⁺CD206⁺, and Tim4⁺CD206^hi KC, and TIM4⁻ TAMs in tumour free and tumour bearing livers from CD45.2 parabionts in (h). j- Gating strategy for separation of Tim4⁺CD206⁺ and Tim4⁺CD206^hi liver KC by flow cytometry. k- Bar-plots show the percentage of Tim4⁺, Tim4⁺CD206⁺, Tim4⁺CD206^hi KC, and TIM4⁻ TAMs labelled with GFP in Cxcr4^gfp/+ mice (n = 3/group) and in Cx3cr1^gfp/+ mice (n = 3/group), and labelled with tdT in Cxcr4^CreERT2;R26^LSL-tdTomato mice pulsed with 4OH-TAM at 6 week-old, that have received intra-portal injection of 1 × 10⁶ KPC-1-luci-tdT cells 2 weeks before analysis (n = 4), or not injected with tumour cells (n = 3/group), not injected with 4OH-TAM (n = 3/group) as control. l- Table represents the percentage (mean and sd) of Tim4⁺ KC, Tim4⁺ CD206⁺ KC, Tim4⁺CD206^hi KC, and TIM4⁻ TAMs in the liver of tumour free and tumour bearing Cxcr4^gfp/+ mice, Cx3cr1^gfp/+ mice, and Cxcr4^CreERT2;R26^LSL-tdTomato mice pulsed at 6 weeks with 4OH-TAM. m- Immunofluorescence staining for F4/80, Tim4, GFP and tdTomato on frozen liver section from Cx3cr1^gfp/+ tumour bearing mice in (k). n- immunofluorescence staining for F4/80, Tim4 and tdTomato on frozen liver section from Cxcr4^CreERT2 R26^LSL-tdTomato tumour bearing mice in (k). o- Cxcr4^CreERT2R26^LSL-tdTomato mice are pulsed with 4OH-TAM (n = 3) or PBS (n = 2) at 6 weeks and analysed 2 weeks later. Bar graphs represent the % of tdT⁺ cells determined by flow cytometry among the indicated cell types. Statistics: One-way ANOVA (h,k). Dots represent individual mice. mean ± sd. ns, not significant.

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