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. 2023 Jun 26;14:1173544. doi: 10.3389/fimmu.2023.1173544

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Copyright © 2023 Steffan, Calise, Park, Niu, Yang, Hammock, Jones, Steele and Keller

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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8-HODE is an agonist and 5,8-diHODE is a partial antagonist of G2A. Using the PathHunter β-arrestin assay (DiscoverX, Eurofins) to assess changes in luminescence we screened 8-HODE (A) and 5,8-diHODE (B) at 1, 10, and 100 μM for agonist (grey bars) and antagonist (pink bars) activity. Data was normalized by DiscoverX to the maximal and minimal response observed in the presence of control ligand ((±)9-HODE) and vehicle (DMSO) respectively. For antagonist assays, the data was normalized by DiscoverX to the maximal and minimal response observed in the presence of the (±)9-HODE at its EC80 concentration (16.7μM) with the vehicle. Samples were run as duplicates. Partial activity was characterized by between 20-50 percent efficacy while full activity was characterized by greater than 50 percent efficacy ( Supplemental Table 3 ).