Extended Data Fig. 7. TA-exposed Adgrl3 does not activate the Gαi/o family.
(a) Gβγ release assay testing D2R activation of Gαi1, Gαi2, Gαi3, GαoA and GαoB in HEK full G protein KO cells. In comparison to the HEKΔ7 CRISPR knockout, this cell line also lacks the full Gαi/o family. Luminescence was read 10 min after stimulation with 10 jliM quinpirole. (b) Gβγ release assay testing the T923S/ΔN924-CTF, PAR1-CTF, and PAR1 activation of Gαi1, Gαi2, Gαi3, GαoA and GαoB in HEK full G protein KO cells. Luminescence was read 10 min after stimulation with 1 μM thrombin. All data are normalized to buffer controls and show the BRET effect induced by ligands. Bars show mean ±SEM from 3 independent experimental replicates. One-way ANOVA with Dunnett’s multiple-comparison post-hoc test was performed for each cDNA construct individually, (no receptor (empty vector), T923S/ΔN924-CTF, PAR1-CTF, and PAR1) to determine statistical significance between the No Gα control and each Gα subtype (For Gαi3**p=0.0064, for GαoB **p=0.0032). See Supplementary Data for the full set of p-values.