Figure 6.

Effect of Sper/NO on IRP1 activities and expression in RAW 264.7 cells. Cells were treated with 100 μM Sper/NO for different times. (A) Aconitase activity in cytosolic extracts was measured by spectrophotometry. (B) IRP1–IRE binding activity was analyzed by EMSA in the presence or absence of 2% 2-ME, and the radioactivity associated with IRP1–IRE complexes was quantified by PhosphorImaging. IRP1 activity is expressed as the percentage of the value obtained after exposure to 2% 2-ME. (C) IRP1 levels were analyzed by Western blotting and quantified by densitometry. ◊, Control; ■, Sper/NO.