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. 2019 Feb 27;9:2953. doi: 10.1038/s41598-019-39007-6

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© The Author(s) 2019

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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Characterization of distinct macrophage subsets in APAP-induced liver injury. C57BL/6 mice were injected intraperitoneally with APAP at 400 mg/kg (body weight) to induce acute liver injury. (A) Flow cytometric analysis of hepatic leukocytes at 48 h after APAP challenge. Liver non-parenchymal cells were identified by first gating for live CD45⁺ leukocytes. Kupffer cells were identified as F4/80^hiCD11b^lo. Monocyte-derived macrophages were identified as F4/80^loCD11b^hi. Distinct subsets of monocyte-derived macrophages and neutrophils were identified based on differential Ly6C expression and the FSC/SSC profile. (B) Flow cytometric analysis of the indicated populations as gated in (A). The shaded histograms represent the unstained controls. (C) The percentage and absolute numbers of Ly6C^hiCX₃CR1^lo and Ly6C^loCX₃CR1^hi monocyte-derived macrophages in the livers at each time point were determined by flow cytometry. Data shown are representative of three independent experiments (n = 3/group).