Extended Data Fig. 7: NanoBiT-G-protein-dissociation assay.
From: Insights into G-protein coupling preference from cryo-EM structures of Gq-bound PTH1R
a. The Gq and Gs coupling activity levels titrated by transfection plasmid volumes of the wild-type PTH1R. b, c. Gq- and Gs-coupling activities and expression levels of the PTH1R mutants. PTH-induced Gq and Gs responses were analysed by a NanoBiT-G-protein-dissociation assay (b). Data are shown as logarithms of relative intrinsic activities (Log RAi; see the Methods). Statistical analysis was performed with two-way ANOVA, followed by Sidak’s post-hoc test (ns, p > 0.05; *, p < 0.01; ***, p < 0.001). The exact p-values are provided in source data file. Cell-surface expression levels were measured by flow cytometry analysis (c). Bars and error bars represent mean and SEM, respectively, from 3 independent experiments (b,c). d. The expression levels of the PTH1R mutants. Cell-surface expression levels were measured by flow cytometry analysis. Bars and error bars represent mean and SEM, respectively, from 3–6 independent experiments. The exact sample sizes are provided in Supplementary Tables 3 (a,d).
