Fig. 1.

Receptor Protein and mRNA Expression and Agonist-Stimulated cAMP Production in 293-R2β Cells

A, Cell lysates from untransfected HEK293 or HEK293 cells stably expressing CRH-R2β receptors were fractionated on SDS-PAGE and subjected to immunoblotting with a specific CRH-R2 antibody in the presence or absence of blocking peptide as described in Materials and Methods. Identical results were obtained from three independent experiments. Inset, RT-PCR of CRH-R2 mRNA present in HEK293 cells using specific oligunucleotide primers. The size of the amplified DNA fragment was 378 bp. B, Cells were pretreated with various concentrations of CRH, UCN-I, or UCN-II for 15 min. cAMP production was determined by ELISA. Results are expressed as the mean ± sem of three estimations.