Fig. 3

Liver-specific deletion of Fgf4 aggravated liver inflammation by increasing M1 macrophage levels in EAH mice. A Serum ALT- and AST-activity levels of Fgf4^fl/fl and Fgf4^−/− mice in EAH and control groups. B Representative images of liver sections from EAH or control Fgf4^fl/fl and Fgf4^−/− mice stained with H&E. Scale bars, 200 μm or 50 μm. C Changes in relative mRNA-expression levels of inflammatory factors (Il1b, Il6, Tnfa, Il10, and Mcp1) in the livers of EAH mice or the control Fgf4^fl/fl and Fgf4^−/− mice. D The quantitative results for F4/80 expression in liver sections from EAH mice or the control Fgf4^fl/fl and Fgf4^−/− mice. E Representative immunofluorescence images of F4/80 expression in liver sections from EAH mice or the control Fgf4^fl/fl and Fgf4^−/− mice. Scale bar, 20 μm. F Heatmap of representative differentially expressed genes related to M1-macrophage and M2-macrophage marker genes in livers of Fgf4^fl/fl EAH mice and Fgf4^−/− EAH mice, based on RNA-seq data. G Changes in relative mRNA-expression levels of genes related to M1-macrophage markers (Nos2 and Cd86) and M2-macrophage markers (Arg1 and Cd206) in the livers of EAH mice or the control Fgf4^fl/fl and Fgf4^−/− mice. H Flow-cytometric analysis of changes in liver macrophages (F4/80 + CD11b + cells), M1 macrophages (CD86 + cells), and M2 macrophages (CD206 + cells), along with quantitation of the results. I WB analyses of changes in NOS2 and ARG1 expression in total liver lysates from EAH mice or the control Fgf4^fl/fl and Fgf4^−/− mice, with quantitation of the results. The WB data were quantified using ImageJ software. n = 3–6 mice/group. Statistical comparisons were made using one-way ANOVA with Tukey’s post-hoc test; *p < 0.05, **p < 0.01, ns, not significant