Hippocampal TSPO Expression Analysis

The western blot analysis was conducted as described previously (Wang et al., 2016 (link)). Briefly, hippocampal tissues (3 mm in diameter around both injection sites) were removed and then extracted by RIPA lysis buffer (Applygen, China). Fifty microgram of protein were separated by SDS-PAGE, measured and analyzed by western blot with primary antibody rabbit anti-TSPO (1:1000; Abcam, Cambridge, MA, United States) and β-actin (1:3000; Santa Cruz, CA, United States). The expression of protein was measured by Gel-Pro Analyzer software, Version 3.1 (Media Cybernetics, Rockville, MD, United States) and the TSPO expression was normalized to β-actin. Every experiment was independently repeated no less than four times.