Figure 3.

Signaling Pathways Involved in GR Phosphorylation

U2OS-hGR cells were pretreated for 1 h with indicated compounds at the concentrations found in Table 1, and GR phosphorylation at S203, S211, and S226 was determined after a 1-h Dex treatment. Equal amounts of protein from each treatment were analyzed by immunoblotting with phospho-S226, phospho-S211, phospho-S203, or total GR antibodies. B, Subcellular distribution of GR upon treatment with BAPTA-AM, curcumin, and DRB. U2OS-hGR cells, treated exactly as above, were fixed, and the subcellular location of GR was examined by indirect immunofluorescence using a total GR antibody. The data shown are from a single experiment that is representative of at least three independent experiments. DAPI, 4′,6-Diamidino-2-phenylindole.