Fig. 10.

IP production for WT hGnRHR and various mutants in response to agonist after rescue with pharmacoperone In3. COS-7 cells were transiently transfected with 20 ng of WT or mutant cDNA plus 80 ng of empty vector as described under Materials and Methods. A, mutants were rescued with pharmacoperone (In3) as described under Materials and Methods; In3 was then washed out, and IP production was measured in response to the indicated dose of buserelin. B, WT GnRHR or mutants were incubated in media alone or rescued with pharmacoperone In3 as described under Materials and Methods; In3 was then washed out, and IP production was measured in response to medium alone or agonist (10⁻⁷ M buserelin). C, mutants were rescued with pharmacoperone (In3) as described under Materials and Methods; In3 was then washed out, and IP production was measured in response to antagonist (acyline) dose-response curve containing agonist (10⁻⁹ M buserelin). In all figures, Means ± S.E.M.s are shown for three independent experiments, each performed in replicates of four.