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. Author manuscript; available in PMC: 2018 Jan 8.
Published in final edited form as: Cell Rep. 2017 Jul 5;20(1):224–235. doi: 10.1016/j.celrep.2017.05.070

Figure 3. Cysteine S-Glutathionylation Protects IL-1β from H2O2-Induced Deactivation.

Figure 3

(A) The effect of glutathionylation on H2O2-induced irreversible oxidation of IL-1β assessed by MS. IL-1β was treated with or without GSSG (250 μM) and then H2O2 (100 μM) for another 30 min at 37°C. The cysteine modification was detected by MS, and the relative amount of each cysteine modification was assessed using Skyline software. Representative plots of precursor, precursor (M+1), and precursor (M+2) intensities for each indicated modification are shown. The molecular weights (MW) of peptides containing the indicated modified cysteine are shown.

(B) The effect of glutathionylation on H2O2-induced deactivation of IL-1β assessed by the IκB degradation assay. Indicated amounts of IL-1β were pretreated overnight with GSSG (250 μM) and then H2O2 (100 μM) for another 30 min at 37°C before adding to HeLa cell cultures. Representative immunoblots for IκBα are shown.

(C) Densitometry results. Shown are the means (±SD) of three independent experiments.