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. Author manuscript; available in PMC: 2020 Nov 4.
Published in final edited form as: Nat Immunol. 2020 May 4;21(7):736–745. doi: 10.1038/s41590-020-0669-6

Extended Data Fig. 4. Disulfiram covalently modifies human GSDMD on Cys 191.

Extended Data Fig. 4

(a) Disulfiram was preincubated for 1 hr with N-acetylcysteine (NAC, 500 μM) or medium before evaluating whether it inhibited pyroptosis of LPS + nigericin treated THP-1 cells. Disulfiram 2-fold dilutions ranged from 5–40 μM. Graphs show mean ± s.d; data are representative of three independent experiments with replicates (n= 3) and similar results. Data were analyzed using two-tailed Student’s t-test. Graphs show the mean ± s.d. and data shown are representative of three independent experiments. **P < 0.01. (b,c) nano-LC-MS/MS spectrum for the peptide containing C191 in human GSDMD. Data are representative of three independent experiments. (b) MS/MS spectrum for peptide FSLPGATCLQGEGQGHLSQK modified on cysteine (red) by carbamidomethyl. Protein coverage was 73%. (c) MS/MS spectrum for peptide FSLPGATCLQGEGQGHLSQK modified on cysteine (red) by disulfiram. Protein coverage was 72%.