For the immunohistochemistry, the sections were degreased with paraffin and covered with an antigen retrieval solution at 95∼100 °C, then, blocked with 5% goat blocking serum for 30 min. Primary antibodies Anti-TNF-α (1:2000, Servicebio), Anti-IL-1β (1:1000, Servicebio), Anti-IL-10 antibody (1:1000, Servicebio), Anti-Bax (1:1500, Servicebio), Anti-BCL-2 (1:1500, Servicebio), Anti-Cyt-c (Cytochrome-c, 1:1000, Servicebio), Anti-CHOP (C/EBP homologous protein, 1:2000, Abdonal), Anti-IRE1-α (Inositol-requiring enzyme-1 alpha, 1:2000, Abdonal) and Anti-XBP1 (X-box binding protein 1, 1:2000, Abdonal) were used. Secondary antibodies goat anti-mouse IgG (1:200, Gibco) and the DAB substrate kit (Servicebio) were used. The slices were imaged by using an Olympus upright microscope (Olympus, DP71, Japan).
For immunofluorescence staining, the sections were permeabilized with 0.2% Triton X-100, 10% normal goat serum and 0.1% BSA in PBS for 1 h. Primary antibodies Anti-CD68 (1:1000, Servicebio), Anti-HO-1 (Heme oxygenase-1, 1:1000, Servicebio), Anti-GRP78 (glucose-regulated protein78, 1:1000, Servicebio) antibody were used. TRITC-labeled and FITC-labeled secondary antibody goat anti-mouse IgG (1:200, Gibco) were used. The sections were imaged by using an Olympus inverted fluorescent microscope (Olympus DP72, Japan).
For immunofluorescence staining, the sections were permeabilized with 0.2% Triton X-100, 10% normal goat serum and 0.1% BSA in PBS for 1 h. Primary antibodies Anti-CD68 (1:1000, Servicebio), Anti-HO-1 (Heme oxygenase-1, 1:1000, Servicebio), Anti-GRP78 (glucose-regulated protein78, 1:1000, Servicebio) antibody were used. TRITC-labeled and FITC-labeled secondary antibody goat anti-mouse IgG (1:200, Gibco) were used. The sections were imaged by using an Olympus inverted fluorescent microscope (Olympus DP72, Japan).