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. 2019 Dec 10;8(24):e013465. doi: 10.1161/JAHA.119.013465

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© 2019 The Authors and AstraZeneca. Published on behalf of the American Heart Association, Inc., by Wiley.

This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

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In vitro specificity in relaxin/insulin‐like family peptide receptor 2 (RXFP2) (A), RXFP3 (B), RXFP4 (C), or ADCYAP1R1 (D) CHO cells. The in vitro specificity of RELAX10 was tested in human RXFP2‐, RXFP3‐, RXFP4‐, or ADCYAP1R1‐expressing CHO cells for the stimulation or inhibition of cAMP production; relative light units (RLUs) represent the cAMP concentration. cAMP assays were performed as described for RXFP1 cell line, except for cell lines RXFP3 and RXFP4, in which 25 or 20 μmol/L forskolin (R&D Systems), respectively, was added to the dilution plate and serum‐free medium to increase intracellular levels of cAMP as the ligands used in the assay will inhibit cAMP production. INSL3 (RXFP2), relaxin‐3 (RXFP3), INSL5 (RXFP4), and PACAP1‐27 (ADCYAP1R1) were used as positive controls. Experiments were performed at least twice.