Anti glp 1r

The tissues were vacuum inflation-fixed and paraffin embedded, and 5-μm sections were cut. Sections were used for immunohistochemical examination. Immunohistochemical staining was performed with the following primary antibodies: rabbit monoclonal anti-LC3 (Cell Signaling Technology), rabbit recombinant monoclonal anti-p62 (Bimake), rabbit monoclonal anti-FN (Abcam), and rabbit polyclonal anti-GLP-1R (Bioss) followed by incubation with horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG secondary antibody according to the kit instructions (ZSGB-BIO, Beijing, China). Peroxidase conjugates were subsequently visualized by using diaminobenzidine (DAB) solution. The sections were then counterstained with hematoxylin and mounted on cover slips. Between each step, the cells were extensively rinsed three times for 5 min each time. HE and Masson staining were performed according to the manufacturer’s instructions. The staining kits were purchased from Nanjing JianCheng Bioengineering Institute (Nanjing, China). Staining was photographed using a Leica microscope (DM2500, Wetzlar, Hesse, Germany). For each rat, five fields were chosen to obtain the average of integrated optical density (iod) of IHC staining and the average collagen area indicated by Masson staining by Image Pro Plus (IPP) software. Average areas from 5 rats were determined, and the analysis was done blindly.

Western blot analysis was performed as described previously. [35 (link)] Primary antibodies used in this study were anti-GLP-1R (Bioss) and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH; Sigma-Aldrich). The intensity of each band was determined using ImageJ software.