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note
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2. To evaluate cell viability, MTS [3-(4,5 dimethylthiazol-2-yl) 5-(3-carboxymethoxy-phenyl)-2-(4- sulphonyl)-2H-tetrazolium, inner salt] (2.0 mg/ml) and PMS (Phenazine methosulfate) (0.92 mg/ml) were added in a ratio of 10:1 (20 μl per well) and plates incubated for 3 h at 37 °C. Resultant absorbance was measured at 490 nm in an ELISA reader. The specific absorbance that represented formazan production was calculated by substraction of background absorbance from total absorbance. The mean % viability was calculated as follows: Mean specific absorbance of treated cells Mean specific absorbance of untreated cells × 100
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79955561622
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note
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2, pH 7.4). Annexin V-FITC and propidium iodide were then added and incubated for 15 min in dark at 25 °C according to the manufacturer's instructions. Data acquisition was done on a FACS Calibur flow cytometer (BD Biosciences, USA) and analyzed with cell quest pro software.
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70350525419
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For flow cytometric analysis of externalized phosphatidylserine, see: P. Saha, R. Sen, C. Hariharan, D. Kumar, P. Das, and M. Chaterjee Free Radical Res. 43 2009 1101
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