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. 2019 Feb 12;9:1861. doi: 10.1038/s41598-019-38539-1

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as 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 images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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The effects of activators of GPR84 in RAW264.7 cells are on-target. Membranes were generated from LPS-treated RAW264.7 cells (a,b). These were used in [³⁵S]GTPγS binding studies in which the effect of varying concentrations of compound 107 was used to block effects of EC₈₀ concentrations of either 2-HTP (a) or PSB-16671 (b). The ability of compound 107 to block effects of either 2-HTP (c) or PSB-16671 (d) to promote binding of [³⁵S]GTPγS to either human or mouse GPR84-Gα_i2 fusion proteins expressed in Flp-In T-REx 293 cells is also displayed. Data represent means ± S.E.M. of combined data from experiments performed on 7 (a) or 5 (b–d) individual membrane preparations.