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. 2017 Aug 11;7:7885. doi: 10.1038/s41598-017-07392-5

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© The Author(s) 2017

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 subunit expression level impacts on agonist-mediated G protein activation. (a,b) BRET (black) in HEK293T cells co-expressing HA-β1-AR (left panels) or HA-β2-AR (right panels) or (c) AT1a-R receptors and different expression levels of Gαs-Rluc8 (a) or GαoA-Rluc8 (b) or Gα11-Rluc8 (c) in the presence of fixed GFP10-Gγ2 and Gβ3 untagged subunits. Cells were stimulated or not for 1 min with 10 µM of the indicated selective agonists isoproterenol (ISO) or angiotensin II (AngII). Results are expressed as the difference in BRET signals measured in presence and absence of ligand. Data represent the mean ± s.e.m. of at least three independent experiments. The statistical significance between stimulated and unstimulated cells was assessed using paired Student’s t-test (*P < 0.05, **P < 0.01, ***P < 0.001). Gα-Rluc8 relative expression levels (gray) were depicted by the luminescence measurement and represent the mean ± s.e.m. of at least three independent experiments. The statistical significance of the difference in luminescence between Gα expression levels was assessed using one-way ANOVA followed by a Tukey’s multiple comparison test (^$ P < 0.05, ^$$ P < 0.01, ^$$$ P < 0.001).