Fig 1: Glucocorticoid stimulation of cAMP depends upon Gas expression. A, HASM were transfected with siRNA specific for GNAS or scrambled control for 48 hours and lysates analyzed by SDS-PAGE and immunoblotting simultaneously with antibodies specific for Gas and ß-actin. Image is representative of n = 3 experiments on separate donor cells. RNA sequencing revealed GNAS transcript was reduced 6.07 ± 0.35 fold (n = 6) following transfection with siRNA. B-E, cADDis sensor was expressed in control or Gas-knockdown HASM using a recombinant BacMam virus then responses to vehicle, forskolin (C), formoterol (C), budesonide (D), or fluticasone (E) were measured. Each point represents the mean ± SEM of n = 4-5 donors. * denotes P < .05, ** denotes P < .01 of each time point