Anti mhc class 2

The midbrain was placed in a lysis buffer containing protease inhibitors and homogenized by ultrasound on ice, centrifuged at low temperature and the supernatant was collected. BCA quantitative method was used for protein quantification. After protein quantification, the loading buffer was added to each sample and heated in a water bath at 100°C for 5 minutes to prepare the sample. The sample was separated by 10% Bis-Tris gel electrophoresis, transferred to PVDF membrane, blocked in 5% skim milk powder (w/v) for 1 h, and then incubated with anti-Tyrosine Hydroxylase (Abcam, 1:1000), anti-MHC class II (Santa Cruz, 1:500), anti-Arginase-1 antibody (Cell Signaling Technology, 1:1000) at 4°C overnight. The sample was incubated with goat anti-rabbit IgG or goat anti-mouse IgG (Beyotime, 1:1000) at room temperature for 1 h. Blots were imaged by the ECL chemiluminescence (Millipore, USA) and a Gel Image System (GE, USA). Densitometry was performed by using Image J software.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and 3,4-Dihydroxyphenylacetic acid (DOPAC) were provided by Sigma-Aldrich (Saint Louis, USA). Dopamine (DA) and Homovanillic acid (HVA) were provided by Solarbio (Beijing, China). Anti-Tyrosine Hydroxylase antibody was provided by Abcam (Cambridge, UK), anti-MHC class II was provided by Santa Cruz (Dallas, USA), anti-Arginase-1 was purchased from Cell Signaling Technology (Danvers, MA, USA), goat anti-rabbit IgG and goat anti-mouse IgG conjugated to HRP were purchased from Beyotime Institute of Biotechnology (Shanghai, China), and ELISA Kits were provided by R&D (Minneapolis, USA).