Inhibition of NF-κB activity in MM1R cells expressing GR mutants, GR N454D/A458T, and GR C476W/R479Q. (A) Inhibition of NF-κB activity in cells infected with adenoviral vector expressing Iκ superrepressor. Cells expressing mutant GR N454D/A458T and C476W/R479Q were infected with either a control adenovirus (CMV) with no transgene or adenovirus expressing Iκ superrepressor at an MOI of 100 and 25, and 24 hours later transfected with NF-κB luciferase reporter, rel A expression plasmid (PLG033-P65-SP10), and TK-Renilla luciferase reporter (“Reporter assays”). Twelve hours after transfection, cells were analyzed by a dual luciferase assay. Data shown as ratio of firefly to Renilla luciferase, relative to the cells not infected with viral vector. (B) Cells expressing mutant GR N454D/A458T and C476W/R479Q were infected with either a control adenovirus (CMV) with no transgene or adenovirus expressing Iκ superrepressor at MOI 5, 25, and 100 and simultaneously treated with 10−6 M dexamethasone. Caspase assay was performed at 48 hours after infection and initiation of dexamethasone treatment.
