Cells were washed with 1 × PBS and lysed with RIPA buffer (150 mM NaCl, 50 mM Tris pH 8.0, 1% Triton X-100, 0.5% sodium deoxycholate and 0.1% SDS) containing protease and phosphatase inhibitors (Roche, Basel, Switzerland). For in vivo samples, the tissues were homogenized and lysed with RIPA buffer supplemented with protease and phosphatase inhibitors. The protein concentration was measured using Bio-Rad Protein Assay reagent (Bio-Rad Laboratories, CA, USA). Standard Western blotting was performed and the blots were visualized using the chemiluminescence UVP BioSpectrum 600 Imaging System (Fisher Scientific, MA, USA). Antibodies used for immunoblotting include FoxO1 (Cell Signaling, Cat.No. 2880, 1:2,000), FoxO1 (Abcam, Cat.No. ab39670, 1:2,000), p-Creb (Ser133) (Cell Signaling, Cat.No. 9196, 1:2,000), p38 MAPK (Cell Signaling, Cat.No. 9212, 1:2,000), p-p38 MAPK (Cell Signaling, Cat.No. 9211, 1:2,000), anti-TH (Milipore, Cat.No. AB152, 1:1,000), anti-TH phosphoSer40 (Milipore, Cat.No. AB5935, 1:1,000), anti-Dopamine D2 Receptor (Milipore, Cat.No. AB5084P, 1:1,000), c-myc (Roche, Cat.No. 11667149001, 1:1,000), GAPDH (Santa Cruz, Cat.No. sc-25778, 1:10,000), anti-Prolactin (Santa Cruz, Cat.No. sc-7805, 1:500), anti-PGC1α (Abcam, Cat.No. ab54481, 1:2,000), anti-Ucp1 (Abcam, Cat.No. ab10983, 1:10,000), anti-COMT (Abcam, Cat.No. ab126618, 1:1,000), anti-MAO-b (Abcam, Cat.No. ab125010, 1:2,000) and β-Actin (Abcam, Cat.No. ab6276, 1:10,000).
