Figure 6.

Effect of cycloheximide (CHX) on IFN-stimulated transcription factors in RAW cells. (A) Binding of GIFs to GATE. Treatments were indicated on the top of the figure. B was identical to A except that ISRE probe was employed. Position of the IRF-1 was indicated with an arrow. (C) Effect of protein kinase inhibitors on GIF activation. RAW cells were incubated with various protein kinase inhibitors before IFN-γ-treatment, and EMSA was performed with GATE. Genistein (100 μg/ml) and herbimycin (Herb; 3 μM) were added to the cells at least 10 h before IFN-γ (1000 units/ml) treatment for 6 h. Other inhibitors, 10 mM 2-amino purine (2-AP) and 0.5 μM staurosporine, were added 30 min before incubation with IFN-γ. EMSA with GATE probe was performed as above. Positions of GIF complexes were indicated.