Science Advances

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• Fig. S1. HSA persulfide is reduced to HSA by sulfide in a dose-dependent manner.
• Fig. S2. Selenite-reducing TrxR1 activity is not inhibited by polysulfides.
• Fig. S3. Compromised polysulfide reduction by TrxR1 mutants lacking the Sec residue.
• Fig. S4. Detection of persulfide formation on BSA.
• Fig. S5. BSA-SSH reduction by NADPH/TrxR1-coupled Trx1 and TRP14.
• Fig. S6. Relative efficiency of alkylation and streptavidin pulldown in the ProPerDP method applied to control and TrxR1 or TRP14 knockdown HEK293 cells.
• Fig. S7. Control experiment to the BSA-SSH reduction by the NADPH/GR/GSH-coupled Grx1 experiment (see Fig. 6 in the main text).
• Fig. S8. Endogenously detected protein persulfides exhibit elevated levels in HS_x⁻-treated TrxR1, TRP14 knockdown, or control HEK293 cells.
• Fig. S9. Diagonal gel electrophoresis experiment corroborates that false-positive hits outlined in Fig. 8B by ProPerDP are of minor importance.
• Fig. S10. Suggested scheme for polysulfide reduction by the NADPH/TrxR1-coupled TRP14 system.

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