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. 2018 Oct 10;8:15076. doi: 10.1038/s41598-018-33440-9

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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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CX₃CR1 expression assigns two subsets and pro-inflammatory phenotype to F4/80^high or Ly6C^low macrophages in the liver. (a) Liver monocytes and macrophages were analyzed by flow cytometry using isolated liver MNCs of CX₃CR1^+/GFP and CX₃CR1^GFP/GFP mice and 4-week after clodronate liposome treatment. (b) Isolated whole F4/80^high cells were subjected to qRT-PCR analysis at week 4 after clodronate liposome treatment. (c) To generate chimeric mice, recipient wild type mice (CD45.1) were transplanted with donor bone marrow cells (BMC) of CX₃CR1^+/GFP or CX₃CR1^GFP/GFP mice (CD45.2) after irradiation and clodronate liposome treatment. Chimerism was assessed by expression of GFP (CD45.2) in liver MNCs of recipient mice. (d) Isolated liver MNCs from chimeric mice were subjected to flow cytometry. (e) F4/80^highGFP⁺ and F4/80^highGFP⁻ cells of chimeric mice were subjected to qRT-PCR analysis. Data are expressed as the mean ± SEM. *p < 0.05, **p < 0.01 compared to GFP⁻ F4/80^high cells. The results represent two independent experiments.