Fig. 5.

Effect of endogeneous NO production on iNOS activity is rescued by L-NAME and C152S GAPDH: (A) Activated RAW264.7 cells were harvested every 2 h during the 0- to 20-h time-course and analyzed for SNO-GAPDH content by biotin-switch followed by pull-downs with neutravidin-linked Sepharose. Eluates were probed for GAPDH. Western analyses show the levels of iNOS and GAPDH proteins present in the cell supernatant samples. (B) Inducible NOS activity in the cell supernatant samples collected every 2 h during 0–20 h of postinduction (LPS and IFN-γ treatment) in RAW264.7 cells (± L-NAME), GAPDH, or mutant overexpressing stable cell lines.