MC65 Cell culture

MC65 cells were cultured in MEMα supplemented with 10% fetal bovine serum (FBS; GIBCO/Life Technologies), 2 mM L-glutamine (Sigma-Aldrich), and 0.1% tetracycline (Sigma-Aldrich). For each experiment, cells were trypsinized and resuspended in Opti-MEM without phenol red (GIBCO/Life Technologies), then treated with STX or matched DMSO concentrations in the presence and absence of tetracycline. All endpoints were compared to those obtained with tetracycline-treated cells, either with or without the addition of STX. For assays of viability, cells were plated at 10,000 cells/well in 96 well plates and assessed after 72 hr of continuous treatment. For assays of gene expression and ATP determination, cells were plated at 60,000 cells/well in 12 well plates and were harvested after 48 hr of continuous treatment.