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. 2020 May 29;10:8779. doi: 10.1038/s41598-020-65636-3

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© The Author(s) 2020

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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Gα_i2-induced activation by the β₁AR and β₂AR. (a) Schematic representation of the Gα_i2-Rluc/Gγ₁-GFP biosensor used to study the Gα_i induced β₁AR and β₂AR signalling. HEK 293 cells were transfected with (b-e) Gα_i2-Rluc, Gγ₁-GFP and untagged Gβ₁, along with (b-c,e) β₁AR or (d-e) β₂AR. (b,d) Kinetics curves represent time course of Gα_i2 activation by (b) β₁AR (vehicle and isoproterenol, n=3) or (d) β₂AR (vehicle and isoproterenol, n=3) expressed as absolute BRET ratios. (c) Concentration-responses curves for Gα_i2 activation following β₁AR activation by indicated ligands. (e) Concentration-responses curves for Gα_i2 activation following isoproterenol-induced activation of β₁AR or β₂AR (n=2). Data were normalized to maximal isoproterenol response, which was take as 100%, and are expressed as mean ± SEM values. Detail (c) of the number of experiments, maximal responses, pEC₅₀ values and statistical comparisons of curve parameters for different ligands are provided in Supplementary Tables S1 and S2.