*4.3. Western Blot Analysis*

Homogenized slice lysates were prepared with lysis buffer (PRO-PREPTM, Intron Biotechnology, Burlington, MA, USA). Proteins were resolved by SDS-PAGE (100 μg/lane) using 10% (*w*/*v*) polyacrylamide gels and transferred to PVDF membranes (Millipore). Membranes were incubated with antibodies against anti-PSD 95 (1:5000, Abcam, Cambridge, MA, USA); anti-synapsin I (1:3000, Abcam); superoxide dismutase (SOD) (1:7500, Abcam); and Nrf2 (1:5000, Abcam) for 2 h at room temperature. As a loading control, membranes were also probed with anti-β-actin antibody (1:10,000, Abcam). The reaction was developed with an enhanced chemiluminescence Western blot analysis system (ECL, GE Healthcare, Marlborough, MA, USA). Signal intensities were analyzed using a gel-scanning integrated optical density software program (Multi-gauge 3.0, Fuji film, Tokyo, Japan).
