Fig. 8.

Knockdown of glucocorticoid receptor-α (GRα) had little effect on budesonide or budesonide + formoterol-induced cAMP production in human airway smooth muscle (HASM). HASM cells were transfected with siRNA specific for GRα or scrambled control (mock) for 72 h. A: lysates were collected and analyzed by immunoblotting using antibodies specific for GRα or β-actin. Each lane shows one of 3 separate experiments on individual cell lines. B, C and D: cAMP difference detector in situ sensor was expressed in HASM with a recombinant BacMam virus, and then cAMP responses to budesonide (1 μM, B), formoterol (0.1 nM, C), or both budesonide and formoterol (1 µM and 0.1 nM, respectively, D) were measured in HASM that was transfected with nontargeting siRNA or siRNA specific for GRα. Each point represents the mean ± SE of n = 4–5 cell lines.