Ab92291

Cells were lysed with RIPA buffer, and 25 μg of protein per lane were separated by 15% SDS-PAGE. Transfer, hybridization, and visualization were made as previously reported [25 (link)]. The primary antibodies were goat polyclonal anti-TSPO (1:1000, abcam, Cambridge, UK, ab92291); rabbit monoclonal anti-galectin 3 (1:1000, abcam, Cambridge, UK, ab76245), and mouse monoclonal anti-actin (1:1000, R&D systems, Minneapolis, MN, USA, MAB8929). The secondary antibodies were anti-goat (Zymed, Waltham, MA, USA, 61-1620), anti-rabbit (Invitrogen, Waltham, MA, USA, G21234), and anti-mouse (Jackson ImmunoResearch, West Grove, PA, USA, 115-035-003).

The rat hippocampus was collected immediately after the animals completed the behavioral tests. Proteins of the tissues were extracted and the concentrations determined by BCA Protein Assay Kit (CWBIO, Beijing, China). Each sample contained with 40 μg of protein was separated by 10 or 12% SDS-PAGE and then transferred to nitrocellulose membranes (Millipore, MA). The following primary antibodies were incubated with the PVDF membrane at 4°C overnight. PBR (1:7,000 dilution, ab92291; Abcam), ATG7 (1:7,000 dilution, ab133528; Abcam), ATG5 (1:7,000 dilution, ab108327; Abcam), LC3B (1:2,000 dilution, ab192890; Abcam), p62 (1:7,000 dilution, ab109012; Abcam) and β-actin (1:7,000 dilution, aa128; Beyotime Biotechnology). After that, rabbit anti-goat immunoglobulin G (IgG) (H + L) horseradish peroxidase (HRP) (1:5,000 dilution, ZB2306; ZSGB-BIO), goat anti-rabbit immunoglobulin G (IgG) (H+L) HRP (1:7,000 dilution, GAR007; MultiSciences) or goat anti-mouse IgG (H+L) HRP (1:7,000 dilution, GAM007; MultiSciences) were incubated with the membrane for 2 h at room temperature. Bands were visualized with enhanced chemiluminescence (ECL) detection reagents (CWBIO, Beijing, China) using an ECL reagent. The relative band intensity was measured by Image-Pro Plus software.