¡fl I |h FeSO4 + SNAP + DTT g QT"^ FeSO4 + SNAP
Caspase 3-like activity [AU/mg protein]
RSNOs [nmol/mg protein]
Fig. 1. Relationship between non-heme iron, iron-nitrosyl complexes, RSNOs and caspase 3 activity in RAW264.7 cells. Cells were precultured with 80 ^M FeSO4 for 24 h, washed with fresh medium and thereafter treated with 0.75 mM SNAP. Cellular levels of iron (A), thiolato-nitrosyl-ironl-2+ complexes (B), and RSNOs (C) were measured as described in Ref. . (B) Spectrum (1) control cells; (2) control cells plus SNAP; (3) iron-loaded cells plus SNAP. (D) RAW264.7 cells were preincubated with FeSO4, washed with fresh medium, and then treated with SNAP. After 15 and 21 h, caspase 3 activity was determined by colorimet-ric assay using the peptide-based substrate Ac-DEVD-p-nitroanilide in the absence and the presence of DTT (10 mM). Results are given as means ± S.D.
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