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2.4. In vitro FFA1 activation assay

AL Alexey Lukin
AB Anna Bakholdina
MC Mikhail Chudinov
OO Oleksandra Onopchenko
EZ Elena Zhuravel
SZ Sergey Zozulya
MG Maxim Gureev
MK Mikhail Krasavin
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CHO cells stably expressing human FFA1 (stable CHO-GPR40 line created at Enamine Ltd.) were seeded (12,500 cells/well) into 384-well black-wall, clear-bottom microtiter plates 24 h prior to assay. Cells were loaded for 1 h with fluorescent calcium dye (Fluo-8 Calcium Assay kit, Abcam, ab112129) and tested using fluorometric imaging plate reader (FLIPR Tetra® High Throughput Cellular Screening System, Molecular Devices Corp.). The maximum change in fluorescence over the base line was used to determine agonist response. A potent and selective agonist for FFA1 GW9508 (Selleckchem, S8014) was tested with the test compounds as a positive control. Concentration-response curve data were fitted using Molecular Devices ScreenWorks® System Control Software (Molecular Devices).

Copyright and License information:  ©2021 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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