Quantifying ROS and NO in Human Endothelial Cells

Estimates of ROS and NO production were made using 2′,7′-dichlorofluorescein diacetate and 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate (diaminofluorescein diacetate; DAF-FM diacetate) (Sigma), respectively, as we described.^{[31 (link)]} To determine insulin-stimulated NO production HAECs were treated with 100 nm insulin for the last 30 min of each respective treatment as we described.^{[24 (link)-26 (link),31 (link),32 (link)]}

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Bharat D., Cavalcanti R.R., Petersen C., Begaye N., Cutler B.R., Costa M.M., Ramos R.K., Ferreira M.R., Li Y., Bharath L.P., Toolson E., Sebahar P., Looper R.E., Jalili T., Rajasekaran N.S., Jia Z., Symons J.D, & Babu P.V. (2017). Blueberry Metabolites Attenuate Lipotoxicity-Induced Endothelial Dysfunction. Molecular nutrition & food research, 62(2), 10.1002/mnfr.201700601.

Publication 2017

DAF-FM diacetate

Daf fm diacetate Insulin

Corresponding Organization : University of Utah

Other organizations : University of Alabama at Birmingham, University of North Carolina at Greensboro

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Variable analysis

independent variables

Insulin (100 nM)

dependent variables

ROS production
NO production

control variables

Treatment duration (30 min)

controls

Positive control: Not specified
Negative control: Not specified

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