Fig. 7.
Effect of cytokine treatment on translational repression activity of IRP in macrophage J774 cells. I-12.CAT (left panel) and I-19.CAT (right panel) mRNA, containing, respectively, the wild-type or mutated ferritin IRE sequence upstream of the CAT open reading frame, were translated in wheat germ extract in the presence of35S methionine. The effect on translational activity of the addition of increasing amounts of lysate (10 to 20 μg protein) from J774 cells untreated (lysate C) or treated with IFN-γ/LPS for 24 hours (lysate IFN-γ/LPS) was evaluated. Translation products were separated on 10% SDS/polyacrylamide gels and visualized by fluorography. The fluorogram shown is typical of four separate experiments.
