Additional file 1: Figure S1. of The antioxidant N-acetyl cysteine suppresses lidocaine-induced intracellular reactive oxygen species production and cell death in neuronal SH-SY5Y cells

Analysis of cell apoptosis by FACS. Levels of cell apoptosis were measured using an Annexin V-FITC Apoptosis Detection Kit (BioVision, Milpitas, CA, USA), according to the manufacturer’s instructions. For these analyses, SH-SY5Y cells were seeded into 6-well plates (3 × 105 cells/well) and incubated overnight. The following day, cells were treated with the indicated concentrations of the appropriate drug(s) for varying lengths of time and harvested by centrifugation at 1200 rpm for 3 min. The culture supernatants were discharged, and the resulting pellets were resuspended in a mixture comprised of 500 μl binding buffer, 5 μl Annexing V-FITC, and 5 μl propidium iodide (PI; 50 μg/ml) for 5 min at room temperature in the dark and analyzed using a FACSCalibur flow cytometer (BD Biosciences, San Jose, CA, USA). (PDF 2009 kb)