Characterization of MM1R cells expressing wild-type GR. (A) Western blot analysis of MM1S, MM1R-EV, and MM1R with wild-type GR-α (MM1R-WT-GR). Two bands in MM1S cells correspond to GR-α and beta isoforms. (B) GRE reporter expression by dexamethasone treatment (10⁻⁶ M for 12 hours) comparing MM1S cells to MM1R cells with empty vector (MM1R-EV) and MM1R-WT-GR. Data shown as fold induction calculated as described in “Reporter assays” and reported as mean plus or minus SD; n = 3. MM1R-WT-GR cells demonstrate significantly increased dexamethasone-induced reporter expression compared with MM1R-EV cells (P < .05). (C) NF-κB transrepression assay, reported as percentage repression of reporter expression on dexamethasone treatment compared with the untreated control (mean ± SD). MM1R-WT-GR cells demonstrate significantly greater NF-κB repression compared with MM1R-EV cells (P < .05). (D) Western blot analysis of MM1R cells for total RAFTK, phospho-RAFTK, and actin with and without dexamethasone treatment (10⁻⁶ M) for 14 hours. (E) Percentage of cells undergoing apoptosis. Caspase 3 assay with flow cytometric analysis of cells treated with 3 concentrations of dexamethasone and analyzed at 60-hour time point. Experiment was done twice with similar results.