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. 2020 Apr 24;27(9):2651–2667. doi: 10.1038/s41418-020-0536-x

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© The Author(s), under exclusive licence to ADMC Associazione Differenziamento e Morte Cellulare 2020

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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Fig. 4 — Liver tissues were collected from mice 6 h after LPS or normal saline (NS) treatment. Also, cultured primary hepatocytes were collected after LPS incubation for 6 h. The following analysises were performed. a Inflammatory foci. H&E staining was performed on paraffin-embedded section of mouse livers. Inflammatory foci were observed and quantified using a microscope. Data are mean ± SD, **P < 0.01 by two-way ANOVA followed by Tukey’s test. n = 3/NS group and n = 6/LPS group. Scale bar = 100 μm (upper panels) and 50 μm (down panels). b Macrophage and neutrophil recruitments. Immunostaining for F4/80 (macrophage) and neutrophils was performed on frozen sections of mouse livers. Hoechst 33342 was counter stained to visualize nuclei. The staining was observed and quantified using a fluorescence microscope. Data are mean ± SD, **P < 0.01 by two-way ANOVA followed by Tukey’s test. n = 3/group. Scale bar = 50 μm. c Expression of mRNA levels. Levels of the indicated mRNA expression in livers (left panel) and primary hepatocytes (right panel) were evaluated using real-time PCR. Data are mean ± SD, *P < 0.05 and **P < 0.01 by Student’s two-tailed unpaired t test. n = 4–11/group.