Synaptosomes were lysed using radioimmunoprecipitation assay (RIPA) buffer [50 mM tris-HCl, 150 mM NaCl, 1% NP-40 (IGEPAL® CA-630, Sigma-Aldrich, catalog #I8896), 0.5% sodium deoxycholate, 0.14 SDS, 1 mM dithiothreitol, and 1X protease inhibitor cocktail] to isolate synaptic proteins. The isolated synaptic protein was quantified using the Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, catalog #23225). Dissociated mouse brains and induced neuronal cultures served as controls and subjected to a similar protocol of protein extraction and lysis using RIPA buffer.
