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. Author manuscript; available in PMC: 2018 Jul 1.
Published in final edited form as: Free Radic Biol Med. 2017 Apr 30;108:741–749. doi: 10.1016/j.freeradbiomed.2017.04.367

Fig. 6.

Fig. 6

Reaction between MNS and glutathione. (A) Michael adduct formation to the enone moiety of MNS followed by a slower rehybridization at C1. (B) Absorbance traces at 378 nm following the addition of 400 μM GSH to 40 μM MNS dissolved in phosphate buffer, pH 7.5. Subsequent addition of 5 mM NEM (50 μL to a final volume of 1.05 mL) reversed the bleaching (see also Fig. S12). The increase in the final absorbance after 2 h compared to the initial reading before GSH addition reflects a small contribution of NEM to the total absorbance at 378 nm. (C) Absorbance changes accompanying the addition of GSH to 40 μM MNS. Data in triplicate are fit to a Kd of 6.7 ± 1.4 μM (see Methods).