Functional characterization of HCA3 orthologs stimulated with aromatic D-amino acids and their metabolites.

CHO-K1 cells were transfected with receptor constructs and cAMP accumulation was determined. The basal cAMP level of mock-transfected CHO-K1 stimulated with 2 μM forskolin (control) in HBSS was 13.8 ± 1.3 nmol/well and set 100%. Emax values are referred to % of mock-transfected CHO-K1 stimulated with 2 μM forskolin in absence of agonist. Emax and EC50 values were determined from concentration-response curves of agonists using GraphPad Prism. A general explanation of EC50 and Emax values is provided in the results section Functional hominoid HCAR analyses with known endogenous agonists. Data is given as mean ± SEM (Emax) and geometric mean with 95% confidence interval (EC50) of at least three independent experiments each performed in triplicates.