Figure 5
Glutamate and VU0424465 trigger mGlu5-dependent Gs and Gq activation. (A) GTPγS fluorescence binding experiments were carried out within detergent purified receptor induced by ligand activation. The fluorescence baseline is estimated based on the receptor alone without any ligands. Data points represent average of three independent measurements with SEM. Dunnett’s test as part of one-way ANOVA was used for comparison with the apo condition set as reference level. (B) mGlu5-dependent Gq activation was measured using the IP one cell-based assay kit (Cisbio) in HEK293 cells and in the presence of VU0424465 with a pEC50 of 8.53 ± 0.13 (n = 4). (C) mGlu5-dependent Gs activation was measured using the cAMP cell-based assay kit (Cisbio) in HEK293 cells and in the presence of glutamate and VU0424465, independently. The dose response curves show a positive response for VU0424465 at the truncated mGlu5-Δ856 (light green, pEC50 of 8.17 ± 0.11; n = 4) and WT human mGlu5 (Dark green, pEC50 of 7.80 ± 0.50; n = 3) curve. A weaker activation is observed for glutamate at the truncated mGlu5-Δ856 (orange, pEC50 of 3.86 ± 0.20; n = 3) and WT human mGlu5 (red, pEC50 of 3.36 ± 0.72; n = 3) curve. Data points represent the average of triplicate measurements from at least three independent experiments with SEM.
