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. 2018 Jul 6;8:10283. doi: 10.1038/s41598-018-28486-8

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© The Author(s) 2018

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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Leukocyte infiltration into HCC in the DEN/TAA/HFD model. (A–C) F4/80 immunostaining (brown) of liver sections, n = 5–6. Scale Bars = 500 μm. (D) Quantitation of F4/80 immunostains (n = 4 mice per group). (E) Clockwise gating strategy from CD45⁺ live cells, showing putative immune cell populations (sub-population number as % of parent population shown in red, remaining cells shown in black). (F–H) Flow cytometry analysis of immune cell sub-populations (n = 3–12 mice per group). KC; Kupffer cell, Neu; neutrophil, Mono; monocyte, pDC; plasmacytoid dendritic cell, NK; natural killer, ILC; innate lymphoid cell, Mac; non-Kupffer macrophages Ly6C^hi; Ly6C^hi monocyte, Ly6C^lo; Ly6C^lo monocyte. Statistical analysis used Kruskal-Wallis test (D), or one-tailed Mann-Whitney test (F–H), *p < 0.05, **p < 0.01.