Figure 6 | Scientific Reports

Figure 6

From: Assessing the combined impact of fatty liver-induced TGF-β1 and LPS-activated macrophages in fibrosis through a novel 3D serial section methodology

Figure 6

(A) Immunohistochemical staining of liver sections. Liver sections were prepared from each group, and then immunohistochemical staining using F4/80 antibody was performed. 3,3′-Diaminobenzidine (DAB) was used to detect the epidermal growth factor module-containing mucin-like hormone receptor-like 1 (F4/80) signals (brown). Nuclei stained with hematoxylin (blue). (B) Serial section-3-dimensional (SS3D) images of the murine livers with macrophage signals. Blood vessel (red), fibrotic tissues (blue), and macrophages (green) were detected and superimposed. Other details of the experiment are included in Materials and Methods. (C) Populations of lymphocyte antigen 6 complex, locus C1 (Ly6C)+ and/or integrin alpha M (CD11b)+ macrophages. To investigate the ratio of Ly6C+ and/or CD11b+ macrophages in total liver blood cells from mice treated as shown in Fig. 1A, fluorescence-activated cell sorting (FACS) analysis was performed with anti-CD11b fluorescein Isothiocyanate (FITC) and Gr-1 (Ly6C) Allophycocyanin (APC) antibodies. Typical data are shown. The other samples were similar in each group [(ND/LPS(−): n = 6; ND/LPS(+): n = 4; HDF/LPS(−): n = 6, HFD/LPS(+): n = 5]. HFD, high-fat diet; ND, normal diet; LPS, lipopolysaccharide.

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