Neurobiotin tracer n 2 aminoethyl biotinamide hydrochloride

Forsythoside B (FB, molecular weight 756.7, 99% purity) was purchased from Shanghai Tongtian Biotechnology Co., Ltd. (Shanghai, China). Antioxidant edaravone or redicava (Simcere) was used as a positive control for ischemia experiments. N-Methyl-d-aspartate (NMDA) receptor antagonist d-2-amino-5-phosphonovalerate (AP5, 50 μmol/L), α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate (AMPA)/kainate glutamate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 μmol/L) were purchased from Sigma–Aldrich. Neurobiotin tracer N-(2-aminoethyl) biotinamide hydrochloride (Vector Laboratories), Alexa Fluor 488 or 647-conjugated streptavidin (Molecular Probes), sodium channel blocker tetrodotoxin (TTX, 500 nmol/L, Baomanbio), γ-aminobutyric acid A (GABA_A) receptor antagonist bicuculline (10 μmol/L, Abcam), and GABA_B receptor antagonist CGP55845 (2 μmol/L, Abcam) were prepared before use, and the purity of each standard compound is no less than 98% by high-performance liquid chromatography analysis. FB and edaravone used in behavioral experiments were diluted in sterile normal saline for intravenous injection.

NMDA receptor antagonist d-2-amino-5-phosphonovalerate (AP5) and α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate (AMPA)/kainate glutamate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) were obtained from Sigma (St. Louis, MO, USA). GABA_A receptor antagonist bicuculline was purchased from Abcam (Cambridge, UK). Neurobiotin™ tracer N-(2-aminoethyl) biotinamide hydrochloride was purchased from Vector Laboratories (Burlingame, CA, USA), and molecular probe Alexa 488-conjugated streptavidin was obtained from Invitrogen (Carlsbad, CA, USA). Neuronal medium NbActiv4 was from Brain Bits (Springfield, IL, USA), animal-free murine IL-4 was purchased from Peprotech (Rocky Hill, NJ, USA).