Fig. 2.

DUOX1 activation promotes protein S-glutathionylation. (A) H292 cells were stimulated with 100 µM ATP for the indicated periods of time, after which cells collected in lysis buffer containing 50 mM NEM. Protein lysates were precipitated with TCA and incubated with DTT to reduce protein mixed disulfides with GSH (PSSG), and GSH was analyzed by HPLC after mBrB derivatization. (B) Analysis of PSSG in unstimulated or ATP-stimulated (100 µM; 15 min) H292-CTL or H292-shDUOX1 cells. (C) PSSG analysis in unstimulated or ATP stimulated MTE cells from wild-type (WT) or DUOX1-deficient (DUOX1-KO) mice. Mean±S.E. from 4 replicates in 2 separate experiments. : p<0.05 compared to unstimulated control; ^#: p<0.05 compared to corresponding treatment of H292-CTL cells.