The obtained liver tissues were immersed in formalin solution for at least 24 h, then paraffin embedded in a multifunctional embedding machine. The tissues were then sliced in a rotary microtome.
For immunofluorescence staining, the tissue sections were dewaxed using xylene and placed in citrate buffer for antigen repair. After endogenous peroxidase was blocked, the tissue sections were incubated with donkey serum followed by an antibody against TREM2 (ab86491, Abcam) and F4/80 (30,325 T, CST). Next, the sections were incubated with the associated secondary antibody (112–605‐062 and 711–545‐152, Jackson) and DAPI. After being sealed with 50% glycerin, the samples were observed and photographed via microscopy (Leica).
For immunohistochemistry experiments, the previously dewaxed and antigen repaired tissue sections were treated using an immunohistochemical kit from DAKO (Denmark). Briefly, samples were treated with peroxidase blocking reagent, an antibody against TREM2 (ab86491, Abcam, USA), Envision FLEX/mouse (linker) reagent, Envision FLEX/HRP reagent, DAB, and haematoxylin in sequence. Finally, the samples were observed and photographed under a microscope (Leica). Dark brown staining areas were considered positive.
For immunofluorescence staining, the tissue sections were dewaxed using xylene and placed in citrate buffer for antigen repair. After endogenous peroxidase was blocked, the tissue sections were incubated with donkey serum followed by an antibody against TREM2 (ab86491, Abcam) and F4/80 (30,325 T, CST). Next, the sections were incubated with the associated secondary antibody (112–605‐062 and 711–545‐152, Jackson) and DAPI. After being sealed with 50% glycerin, the samples were observed and photographed via microscopy (Leica).
For immunohistochemistry experiments, the previously dewaxed and antigen repaired tissue sections were treated using an immunohistochemical kit from DAKO (Denmark). Briefly, samples were treated with peroxidase blocking reagent, an antibody against TREM2 (ab86491, Abcam, USA), Envision FLEX/mouse (linker) reagent, Envision FLEX/HRP reagent, DAB, and haematoxylin in sequence. Finally, the samples were observed and photographed under a microscope (Leica). Dark brown staining areas were considered positive.