Fitc dextran 40000da

Male mice were anesthetized, and fluorescein isothiocyanate dextran (FITC-dextran-40000da, 100 μl, 50 mg/ml, in saline, Sigma-Aldrich, St. Louis, MO) was injected intravenously in the inferior vena cava vein. After 1 minute, the brains were removed and frozen quickly on dry ice. Coronal sections (20 μm thickness) were taken using the cryostat (Leica Microsystems, CM1860, Buffalo Grove, USA). Fluorescent images of the sections were captured using a Nikon Eclipse Ni microscope and NIS Elements software.

GL was purchased from Sigma-Aldrich (Sigma-Aldrich, USA). Gentamycin (GM) was obtained from Amersco (Amersco, USA). LPS (Escherichia coli 0111:B4) and FITC-dextran (40,000 Da) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Recombinant mouse GM-CSF and IL-4 were obtained from PeproTech Inc. (RockyHill, NJ, USA). Anti-mouse antibodies FITC-CD11c, -CD40, -CD80, -CD83 and -CD86, Alexa Fluor®647-TLR2 and PE-MHC-II as well as anti-NF-κB p65 were obtained from Biolegend (San Diego, CA, USA). The ELISA kits for IFN-γ, TNF-α, NO, IL-6, IL-10 and IL-12p70 were obtained from eBioscience (San Diego, CA, USA). Antibodies against β-actin, phospho-JNK, JNK, phospho-p38, p38, IκBα, LaminB1 and HRP-conjugated anti-mouse and anti-goat IgG were purchased from SantaCruz Biotech (Santa Cruz, CA, USA), and phospho-ERK1/2 and anti-ERK1/2 antibodies were obtained from BD Pharmingen (San Jose, CA, USA). The inhibitors BAY 11-7082, SP600125, SB203580 and U0126 were purchased from Beyotime Biotechnology (Haimen, Jiangsu, China).