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Detection of apoptosis by flow cytometry using Annexin V staining
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Early apoptotic cells were detected using the Annexin V-PI Apoptosis Kit (Beckman Coulter, San Diego, CA, USA). Cells were incubated with sTR, FA1FT, HA5FT or HA5ST for 6 h and washed with PBS. The cells were then resuspended in binding buffer, stained with Annexin V (0.6 mg/ml) and propidium iodide (PI, 5 mg/ml) for 15 min in the dark at room temperature and analyzed by two-color flow cytometry (Becton Dickinson, Franklin Lakes, NJ, USA). Data were analyzed using Cell Quest software. Annexin V-FITC fluorescence was detected in FL-1, and PI was detected in FL-3.
Copyright and License information: ©2016 Macmillan Publishers Limited
Cell Death and Disease is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
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