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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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Gα₁₂-induced activation by the β₁AR. (a) Schematic representation of the Gα₁₂-Rluc/Gγ₁-GFP biosensor used to study the Gα₁₂ induced βAR signalling. (b) Schematic representation of the Gα₁₂-Rluc/p115-RhoGef-GFP (p115-GFP) biosensor used to study the Gα₁₂ induced βAR signalling. HEK 293 cells were transfected with (c,e,g) Gα₁₂-Rluc, Gγ₁-GFP and untagged Gβ₁ or with (d,f) Gα₁₂-Rluc, p115-GFP and untagged Gγ₁ and Gβ₁, along with β₁AR or (g) β₂AR. Kinetic curves represent time course of (c) Gα₁₂ activation (vehicle and isoproterenol n = 3) or (d) Gα₁₂-p115 biosensor activation (vehicle n = 2, isoproterenol n = 3), expressed as absolute BRET ratio. Concentration-responses curves for (e) Gα₁₂ activation or (f) Gα₁₂-p115 biosensor activation following β₁AR activation by indicated ligands. (g) Concentration-responses curves for Gα₁₂ activation following isoproterenol-induced β₁AR or β₂AR stimulation (n = 6). Data were normalized to maximal isoproterenol response (100%), and are expressed as mean ± SEM values. Detail (e,f) of the number of experiments, maximal responses, pEC₅₀ values and statistical comparisons of curve parameters for (e,f) β₁AR activation by different ligands are provided in Supplementary Tables S1 and S2.