MTS Assay for Cell Viability

Cell viability was assessed with the MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)2-H-tetrazolium] assay as described previously^{5 (link)}. Cells were seeded into 96-well plate at a density of 3,000/well. Following the OGD and reoxygenation, 20 μl of MTS (5 mg/ml) per well was added, and cells were incubated for additional 2 h. Then, the plates were placed in a microplate autoreader (Thermo, NY, USA), and the absorbance at a wavelength of 490 nm was measured. Cells were seeded and assayed in triplicate for each experimental condition. Cell viability was calculated as the ratio of absorbance in treated cells to that in control cells. Cell growth curves were obtained using GraphPad Prism 6.0.