Hippocampus samples were extracted and homogenized in lysis buffer containing protease and phosphatase inhibitors, and the homogenates were centrifuged at 7000 g for 15 mins at 4°C as described previously (Wu et al. 2012 (link)). The supernatants were collected and the protein concentration was determined by Bradford method, and 30 μg of proteins were loaded for each lane and separated by SDS-PAGE. After the transfer, the blots were incubated overnight at 4°C with one of the primary antibodies listed below: CSF-1 (1:500, R&D, AF416), β-actin (1:5000, rabbit; Santa Cruz). These blots were then washed, incubated with HRP-conjugated secondary antibody and developed in an enhanced chemiluminescent solution. Specific bands were evaluated by apparent molecular size. Finally, images were captured using a Fuji-Film LAS-300 (Fuji, Sheffield, UK) and the intensity of the selected bands was analyzed using Image J software.