SH-SY5Y Cell Culture

SH-SY5Y neuroblastoma cells were cultured in DMEM/F12 medium (GIBCO/Life Technologies) supplemented with 10% FBS and 1% penicillin-streptomycin (Sigma-Aldrich). For assays of gene expression and ATP production, cells were plated at 200,000 cells/well in 12-well plates. For assays of cell viability, cells were plated at 15,000 cells/well in 96 well plates. Three days after plating, cells were washed with phosphate-buffered saline (PBS) and switched to serum-free DMEM/F12 medium containing 1% N-2 growth supplement (GIBCO/Life Technologies) plus STX (100 nM). The following day, the cells were treated with 50 µM Aβ_25–35 (American Peptide Company), a fragment that has been shown to recapitulate the toxic effects of full-length Aβin vitro [42, 43]. Aβ_25–35 solutions were incubated at 37° C for 72 hr, prior to addition to the cell cultures. All endpoints were assessed after 48 hr of treatment.