*4.10. Protein Isolation*

For total protein isolation, tissues obtained from dissected midbrains and striata were homogenized in radio-immunoprecipitation assay (RIPA) buffer (50 nM Tris/HCl, pH 8; 150 nM NaCl; 1% NP-40; 0.5% sodium deoxycholate; 0.1% SDS) and 1x Halt Protease

and Phosphatase Inhibitor Cocktail (Pierce, Thermo Fisher Scientific) with a moto-driven Bio-vortexer (No 1083; Biospec Products, Bartlesfield, UK). Lysates were maintained on ice for 30 min and were then sonicated and centrifuged at 10,000× *g* for 10 min. Supernatants were collected and used as total protein extracts. Protein concentrations were determined by using the Bio-Rad protein assay kit, according to the manufacturer's instructions.
