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bTSH, rhTSH, and thyrostimulin caused inverted U-shaped dose response regulation of cAMP production. HEK-TSHR cells were incubated with increasing doses of bTSH, rhTSH [0–300 mU/ml (5.4 µM)], or thyrostimulin [0–1000 ng/ml (29 nM)] in HBSS/HEPES with IBMX at 37°C for 60 minutes. After incubation with ligands, HEK-TSHR cells were lysed and cAMP was measured in the cell lysates. The symbols represent the mean ± S.E.M. of duplicate measurements in three experiments. Rmax, maximum response.
