Figure 2.

CaSR activation of the G_i/o family by NanoBiT G protein dissociation assays. NanoBiT dissociation assays of AdHEK-CaSR cells transiently transfected with: (A) LgBiT-Gα_i1, (B) LgBiT-Gα_i2, (C) LgBiT-Gα_i3, (D) LgBiT-Gα_o and (E) LgBiT-Gα_z with SmBiT-Gβ subunits (Gβ1–5) and unlabelled Gγ2 following treatment with 5mM Ca²⁺_e. Pre-calcium baselines are not shown. The first baseline value is shown. (F) Percentage suppression of baseline from A, B, C, D and E. Statistical analyses comparing values of all Gα-Gβ dissociations are shown in Supplementary Table 1. For all panels only those Gα and Gβ pairs that yielded relative luminescence units (RLU) over a threshold of 1 × 10³ (i.e. above background luminescence) are shown. All responses were normalised to those under basal conditions (0.1 mM Ca²⁺_e). Dissociation curves for 5 mM Ca²⁺_e are shown (curves with 0.1 mM Ca²⁺_e are shown in Supplementary Figure 8). Curves show mean ± s.e.m. for n = 6–10 independent assays.