Fig. 3.

Rapamycin does not augment cAMP levels over β-adrenergic agonist alone. cAMP levels were measured using the HitHunter cAMP EFC chemiluminescent detection assay (Amersham, Inc.), according to manufacturer’s directions. Briefly, 3T3-L1 adipocytes were serum deprived 20 h and pre-treated without or with rapamycin (R) (20 ng/mL) for 30 min. Cells were then incubated in the absence (DMEM) or presence of isoproterenol (Iso) (1 μM or 10 μM) for 15 min (a) or (10 μM) 30 min (b) in the absence or presence of rapamycin (Iso + R). “No ED” (without enzyme donor) and “PBS” represent negative controls for this assay. Bars with different letters are statistically significant at a level of 0.05