6 ohda

SH‐SY5Y human neuroblastoma cells (ATCC) were cultured in DMEM/F12 medium supplemented with 10% heat‐inactivated fetal bovine serum and 100 μg/mL streptomycin. The cells were maintained at 37°C with 5% CO₂ and the culture medium was refreshed every other day. Passaging of the cells was performed every 2–3 days, and cells at passage 3 were used for subsequent experiments.
To induce neurotoxicity, SH‐SY5Y cells were exposed to 25 μM of 6‐OHDA (Sigma) for 24 h. Prior to 6‐OHDA exposure, SH‐SY5Y cells were transfected with plasmids carrying PCBP1 and empty vectors using Lipofectamine 3000 (Invitrogen) 24 h before the 6‐OHDA treatment. After 36 h, the cell growth of SH‐SY5Y cells was assessed as described below.

Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), 4′,6-diamidino-2-phenylindole (DAPI), penicillin–streptomycin, and 0.25% (w/v) trypsin containing 1 mM ethylenediaminetetraacetic acid were purchased from Invitrogen (Carlsbad, CA), and 6-OHDA, 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH), vitamin C, dimethyl sulfoxide (DMSO), Akt inhibitor IV, MEK inhibitor (PD 98059), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO). Nerve growth factor (NGF) was obtained from R&D Systems (Minneapolis, MN, USA). A total antioxidant capacity assay kit was purchased from Abcam (Cambridge, UK). A lactate dehydrogenase (LDH) cytotoxicity assay kit was purchased from Cayman Chemical (Ann Arbor, MI). A Caspase 3/7 activity detection kit was obtained from Promega (Madison, USA). Antibodies for western blotting were purchased from Cell Signaling Technology (Danvers, MA). All chemicals were dissolved in appropriate solvents and stored at −20°C before use to maintain their chemical stability.