2.2. Protein extraction from tissue

Frozen brain tissue was homogenized by mechanical disruption on ice in TNN buffer (50 mM Tris–HCl, pH 7.5, 150 mM NaCl, 0.5% NP40, 1:200 protease inhibitor cocktail) (Calbiochem, San Diego, CA, USA). Once completely homogenized, samples were centrifuged at 13,000×g at 4 °C for 5 min. The supernatant containing the protein was collected and protein concentrations were determined via the Bradford assay.