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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

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posted on 24.10.2016, 05:00 authored by Akihisa Okamoto, Masahiro Tanaka, Chisato Sumi, Kanako Oku, Munenori Kusunoki, Kenichiro Nishi, Yoshiyuki Matsuo, Keizo Takenaga, Koh Shingu, Kiichi Hirota
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)


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