Fig. 5

IFNγ and NF-κB induce iNOS expression in myeloid cells. a J774 cells were treated with IFNγ, LPS, or both IFNγ and LPS for approximately 18 h, and analyzed for iNOS expression by RT-PCR. β-actin was used as a normalization control. b Cells were treated as in A and then analyzed by real time RT-PCR analysis of iNOS expression with β-action as an internal control. c J774 cells were cultured in the presence of Ruxolitinib for 30 min and then treated with IFNγ and LPS as indicated for 18 h. Total lysates were then prepared and analyzed for STAT1 and pSTAT1 levels by Western blotting analysis. d J774 cells were cultured in the presence of Ruxolitinib for 30 min and then treated with IFNγ and LPS for 18 h. iNOS expression was then analyzed by RT-PCR. e J774 cells were transiently transfected with a control vector or a vector containing the dominant negative IκBα-AA mutant, respectively. Cells were treated with IFNγ and LPS for approximately 18 h, and then analyzed for iNOS expression