Fig. 3.
GRX1 modulates the inhibitory effects of H2O2 on IKK-β and NF-κB. (A Top) Cells were treated with 200 μM H2O2 for 5 min, and S-glutathionylation of IKK-β was assessed as in Fig. 2A. (Middle and Bottom) Control Western blots for HA-IKK-β and HA-GRX1, respectively. (B and C) C10 cells overexpressing HA-GRX1 were exposed to agents as before and evaluated after 5 min for IKK activity (B) or after 15 min for IκB-α levels (C). The level of RelA was measured as a loading control. (D) Cells were cotransfected with 6x κB-tk-luc reporter vector and pcDNA3 (filled bars) or HA-GRX1 (open bars) expression vectors and exposed to 200 μM H2O2 for 5 min before treatment with 10 ng/ml TNF-α for 6 h. Luciferase units were corrected for the amount of protein and expressed as the percentage of pcDNA3-transfected, TNF-α-stimulated luciferase activity.