Fig. 5.

Nonligand modulation of GR. (A) Luciferase assays were performed by using A549 cells that were transfected with pENaC-luc and pSV40-rl and treated with dex and forskolin, as indicated. The relative activity of pENaC-luc with dex treatment was normalized to 100%. (B) Relative mRNA levels of the indicated target genes were determined by using qPCR. Cells were treated as indicated for 24 h. *, P ≤ 0.05. (C) Fluorescence polarization assays were performed by using the indicated compounds as potential competitors to GR binding of a fluorescent control ligand. The value in the absence of competitor drug was normalized to 100%. Results are an average of 2 samples; the standard deviation between paired samples was <7% for all data points. (D) U2OS-GR cells were treated with Hpg (2.5 μM), Ros (10 μM), Acl (200 nM), Ciclo (25 μM), or Para (10 μM) for 24 h and then treated with dex or ethanol for 1 h before fixation. GR distribution was visualized by using indirect immunofluorescence. Drug doses were: Hpg (2.5 μM), Ros (10 μM), Acl (200 nM), Ciclo (25 μM), and Para (10 μM).