Ab211265

The proteins were extracted from the striatum tissues using radioimmunoprecipitation assay (RIPA) lysis buffer (Ukzybiotech, Beijing, China) according to the manufacturer's instructions. The protein concentration in the lysates was evaluated using a BCA Protein Assay Kit. Proteins were then separated on an SDS-PAGE and transferred to a polyvinylidene difluoride membrane. Membranes were blocked with 5% BSA-TBST and then incubated with DA1R antibody (ab20066, 1 : 1000), DA2R antibody (ab85367, 1 : 1000), DA3R antibody (ab42114, 1 : 1000), DA4R antibody (ab20424, 1 : 500), DA5R antibody (ab40656, 1 : 200), PKA antibody (ab211265, 1 : 1000), CAM (ab45689, 1 : 1000), and CAMKII (ab52476, 1 : 1000) (Abcam) overnight at 4°C. Then, the blots were washed, incubated with 5% BSA-TBST, and washed again. Consequently, samples were incubated with horseradish peroxidase-conjugated secondary antibody at room temperature for 2 h. Band detection was performed using the enhanced chemiluminescence (ECL) detection kit. The intensity of the detected bands was calculated densitometrically using the Gel Image system ver.4.00 (Tanon, China).

Total cellular protein was extracted and 25 μg was used for SDS-PAGE. After blocking the membrane for 30 min, rinsing with TBST buffer three times, and incubation with anti-MAP2 (1:1000; Abcam; ab5392), anti-PKA (1:400; Abcam ab211265), and anti-PPAR-γ (1:500; Abcam ab45036) for 12 h at 4 °C, the membranes were washed three times in wash buffer (0.2% gelatin, 0.05% Tween 20 in TBS). The membranes were then incubated for 1 h with horseradish peroxidase-labeled secondary antibody, washed again, and exposed to ECL color development reagents. The membranes were developed using the ChemiDoc-It TM TS2 Imaging System (Bio-Rad), and relative optical density was analyzed using the ImageJ2x software (National Institute of Health, Bethesda, MD, USA).