Figure 2.

Effect of inhibiting protein synthesis or protein trafficking on the ability of IN3 to rescue the receptors. Stably transfected HeLa cells or Cos-7 cells transiently transfected with the E⁹⁰K mutant GnRHR were treated with either the protein synthesis inhibitor cycloheximide (20 μg/ml) or monensin (10 μM) for 30 or 60 minutes prior to addition of 1 μg/ml IN3 for 4 hours (“E”). Inhibitors were present up until the end of the agonist challenge, at which point total IP production was assessed. (A): cells were pre-loaded for 18 hours with ³H-inositol (“G”). (B): cells were pre-loaded for 6 hours with ³H-inositol (“F”). Cells were stimulated with 100 nM Buserelin for 2 hours (“H”). Averages and SEMs were calculated from at least 3 independent experiments performed in replicates of 6.