A high-throughput fluorescence resonance energy transfer (FRET)-based endothelial cell apoptosis assay and its application for screening vascular disrupting agents
In this study, we developed a high-throughput endothelial cell apoptosis assay using a fluorescence resonance energy transfer (FRET)-based biosensor. After exposure to apoptotic inducer UV-irradiation or anticancer drugs such as paclitaxel, the fluorescence of the cells changed from green to blue. W...
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sg-ntu-dr.10356-1004452020-03-07T11:35:25Z A high-throughput fluorescence resonance energy transfer (FRET)-based endothelial cell apoptosis assay and its application for screening vascular disrupting agents Zhu, Xiaoming Fu, Afu Luo, Kathy Qian School of Chemical and Biomedical Engineering In this study, we developed a high-throughput endothelial cell apoptosis assay using a fluorescence resonance energy transfer (FRET)-based biosensor. After exposure to apoptotic inducer UV-irradiation or anticancer drugs such as paclitaxel, the fluorescence of the cells changed from green to blue. We developed this method into a high-throughput assay in 96-well plates by measuring the emission ratio of yellow fluorescent protein (YFP) to cyan fluorescent protein (CFP) to monitor the activation of a key protease, caspase-3, during apoptosis. The Z′ factor for this assay was above 0.5 which indicates that this assay is suitable for a high-throughput analysis. Finally, we applied this functional high-throughput assay for screening vascular disrupting agents (VDA) which could induce endothelial cell apoptosis from our in-house compounds library and dioscin was identified as a hit. As this assay allows real time and sensitive detection of cell apoptosis, it will be a useful tool for monitoring endothelial cell apoptosis in living cell situation and for identifying new VDA candidates via a high-throughput screening. 2013-07-09T02:47:34Z 2019-12-06T20:22:42Z 2013-07-09T02:47:34Z 2019-12-06T20:22:42Z 2012 2012 Journal Article Zhu, X., Fu, A., & Luo, K. Q. (2012). A high-throughput fluorescence resonance energy transfer (FRET)-based endothelial cell apoptosis assay and its application for screening vascular disrupting agents. Biochemical and Biophysical Research Communications, 418(4), 641-646. 0006-291X https://hdl.handle.net/10356/100445 http://hdl.handle.net/10220/11041 10.1016/j.bbrc.2012.01.066 en Biochemical and biophysical research communications © 2013 Elsevier Inc. |
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In this study, we developed a high-throughput endothelial cell apoptosis assay using a fluorescence resonance energy transfer (FRET)-based biosensor. After exposure to apoptotic inducer UV-irradiation or anticancer drugs such as paclitaxel, the fluorescence of the cells changed from green to blue. We developed this method into a high-throughput assay in 96-well plates by measuring the emission ratio of yellow fluorescent protein (YFP) to cyan fluorescent protein (CFP) to monitor the activation of a key protease, caspase-3, during apoptosis. The Z′ factor for this assay was above 0.5 which indicates that this assay is suitable for a high-throughput analysis. Finally, we applied this functional high-throughput assay for screening vascular disrupting agents (VDA) which could induce endothelial cell apoptosis from our in-house compounds library and dioscin was identified as a hit. As this assay allows real time and sensitive detection of cell apoptosis, it will be a useful tool for monitoring endothelial cell apoptosis in living cell situation and for identifying new VDA candidates via a high-throughput screening. |
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School of Chemical and Biomedical Engineering |
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School of Chemical and Biomedical Engineering Zhu, Xiaoming Fu, Afu Luo, Kathy Qian |
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Article |
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Zhu, Xiaoming Fu, Afu Luo, Kathy Qian |
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Zhu, Xiaoming Fu, Afu Luo, Kathy Qian A high-throughput fluorescence resonance energy transfer (FRET)-based endothelial cell apoptosis assay and its application for screening vascular disrupting agents |
author_sort |
Zhu, Xiaoming |
title |
A high-throughput fluorescence resonance energy transfer (FRET)-based endothelial cell apoptosis assay and its application for screening vascular disrupting agents |
title_short |
A high-throughput fluorescence resonance energy transfer (FRET)-based endothelial cell apoptosis assay and its application for screening vascular disrupting agents |
title_full |
A high-throughput fluorescence resonance energy transfer (FRET)-based endothelial cell apoptosis assay and its application for screening vascular disrupting agents |
title_fullStr |
A high-throughput fluorescence resonance energy transfer (FRET)-based endothelial cell apoptosis assay and its application for screening vascular disrupting agents |
title_full_unstemmed |
A high-throughput fluorescence resonance energy transfer (FRET)-based endothelial cell apoptosis assay and its application for screening vascular disrupting agents |
title_sort |
high-throughput fluorescence resonance energy transfer (fret)-based endothelial cell apoptosis assay and its application for screening vascular disrupting agents |
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2013 |
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https://hdl.handle.net/10356/100445 http://hdl.handle.net/10220/11041 |
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1681036155513995264 |