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The 10,000×g cytosolic fractions obtained from COS-7 cells (100 μg of protein) were incubated with 2-arachidonoyl-[3H]-glycerol (40 Ci/mmol, St. Louis, MO, USA) diluted with non-radiolabeled 2-AG (20 μM) in the presence of vehicle or increasing concentrations of PEA (0.1-25 μM), in Tris-HCl 50 mM pH 7.4 at 37 °C for 20 min [42]. After the incubation, the amounts of [3H]-glycerol were measured in the aqueous phase [after extraction of the incubation mixture with 2 volumes of CHCl3/CH3OH (1:1, v/v)] by using Liquid Scintillation Analyzer (TRI-carb 2100TR).
Copyright and License information: The Author(s). ©2019
Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
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