“…During thawing, samples were treated with protease inhibitors (Complet ultra tablet, Roche Diagnostics, Meylan, France) and buffered with NaOH 1 mol/L to adjust the acidic pH of the extraction buffer. Then, equal quantities of proteins were electrophoresed on 12% sodium polyacrylamide-dodecyl sulfate gel with a 4% stacking gel and transferred to nitrocellulose membranes (Amersham 300 × 4 mm, UGAP, Champssur-Marne, France) as previously described [53]. Membranes were saturated 1 h in 5% milk (Regilait, UGAP, Champs-sur-Marne, France) or 5% BSA (P06-139,1100, Dutscher, Montsaunès, France) in Tris-Buffered Saline (TBS) and Tween 0.1%, then incubated with different primary antibodies overnight at 4 • C: anti-DRD1 (D2944; Sigma, Molsheim, France), anti-DRD2, anti-DAT (AB5084P, AB2231 respectively; Millipore, Molsheim, France), anti-SERT, anti-5-HTR1a (ab172884, ab85615 respectively; Abcam, Cambridge, UK) and anti-GAPDH (glyceraldehyde 3-phosphate dehydrogenase) as maintenance protein (5174S, rabbit; Cell Signaling, Leiden, The Netherlands).…”