Fig. 1.
The GPR35 antagonists CID-2745687 and ML-145 blocked agonist-induced hGPR35 activation in 2 distinct assays. Activation of hGPR35 by varying concentrations of the agonists pamoic acid and zaprinast in a β-arrestin-2 interaction assay (a), where FLAG-hGPR35-eYFP was cotransfected with β-arrestin-2-Renilla luciferase in HEK293T cells (n = 6), or an IP1 accumulation assay (b), where FLAG-hGPR35-eYFP was cotransfected with Gαq/135 in HEK293T cells (n = 5). hGPR35 activation following cotransfection of FLAG-hGPR35-eYFP and Gαq/135 in HEK293T cells measured via an IP1 accumulation assay was inhibited in a concentration-dependent manner following exposure to increasing concentrations of ML-145 and CID-2745687 in the presence of either zaprinast (c; n = 5) or pamoic acid (d; n = 7) at EC80 concentrations of the agonist ligand. Data are shown as means ± SEM.