Functional characterization of HCA<sub>1</sub>, HCA<sub>2</sub>, and HCA<sub>3</sub> orthologs.
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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) was 41.1 ± 1.9 nmol/well in DMEM and 13.8 ± 1.3 nmol/well in HBSS and set 100%. Analyses of HCA1 were carried out in DMEM. HCA2 and HCA3 were assayed in HBSS. Emax values are shown as % of mock-transfected CHO-K1 stimulated with 2 μM forskolin in absence of agonist. Emax and EC50 values were determined from concentration-response curves 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 (basal and Emax) and geometric mean with 95% confidence interval (EC50) of at least three independent experiments each performed in triplicates.
