Figure 5.

Modulation of IRP1 expression in C58 pre-T cells cocultured with NO-producing RAW 264.7 cells. RAW 264.7 cells were grown for 16 h in the presence or absence of 10 units/ml IFN-γ and 50 ng/ml LPS. Cells were washed to remove IFN-γ and LPS before addition of C58 cells. C58 cells were cultured for 1–6 h with control (C) and activated (IFN-γ/LPS) RAW 264.7 cells and then were withdrawn from the macrophage monolayer. Nitrite production was measured in the culture medium at each time point. (A) Cytosolic extracts from C58 cells were analyzed for IRP1–IRE binding activity by EMSA. (B) IRP1 expression in C58 cells was determined by Western blot analysis with anti-IRP1 antiserum.