Fig. 2.

Oxidative stress is increased with TAK1 inhibition, and cells respond to TAK1 inhibition with increased intracellular glutathione. Cells were treated with 5 µM 5Z-7-oxozeaenol and then 10 mM N-acetylcysteine. Vehicle=0.1% DMSO. (A) Flow cytometry was used to analyze intracellular reactive oxygen species generation after 24 h of treatment using H₂DCFDA (redox-sensitive probe) and DCFDA (redox-insensitive probe). Data normalized to vehicle control and presented as fold change of mean fluorescence intensity for H₂DCFDA-stained cells/DCFDA-stained cells. (B, C) After 24 h treatment with TAK1 inhibitor, cells were scrape-harvested and reduced glutathione (GSH), (B) and oxidized glutathione (GSSG), (C) levels were assayed. Normalized to vehicle control. Errors represent ±1 SEM of at least 3 separate experiments. *P <0.01, **P <0.05.