Western Blot Analysis of Cortical Proteins

The preparation of cortical tissue extraction for immunoblotting was followed by the method described in our previous study^{63 (link)}. Protein extracts were subjected to 10% Bis-Tris gel (Invitrogen, Grand Island, NY, USA), incubated with 5% non-fat dry milk in TBST buffer (20 mM Tris-HCl, 150 mM NaCl, 0.1% Tween-20) for 1 h at room temperature, and then followed by the primary antibodies with gently shaking overnight at 4 °C. The primary antibodies used were as follows: anti-EP (Cat# 36-3000, Invitrogen), anti-phos-p38 (Cat# 612288, BD), anti-total-p38 (Cat# 9212, Cell signaling), anti-human Aβ 1-17 clone 6E10 (Cat# 9320-02, Signet), anti-synaptophysin (Cat# MAB5258; Chemicon), anti-synaptojanin 1 antibody (AC1), (Cat# MA3-936; Thermo Fisher), anti-NMDAR2B (Cat# ab81271, Abcam), anti-BACE1 antibody (Cat# ab108394, Abcam), and β-actin (Cat# A5441; Sigma-Aldrich). ImageJ software (National Institutes of Health, Bethesda, MD, USA) was used for quantification of intensity of the immunoreactive bands in the scanned blots.