Fig. 5.
Attenuation of NF-κB activation and chemokine production by primary airway epithelial cells from Glrx1−/− mice in response to LPS. GRX1 activity in WT cells was 37.4 units (undetectable in Glrx1−/− cells). (A) Primary tracheal epithelial cells (MTE) were loaded with 1.5 mM Bio-GEE for 1 h, lysates were resolved by nondenaturing SDS/PAGE, and blots were reacted with streptavidin-HRP. IB:β-actin is a loading control. (B) WT or Glrx1−/− MTE cells were treated with 1 μg/ml LPS for 4 h for evaluation of RelA nuclear translocation. Red, RelA immunoreactivity; green, nuclear Sytox green counterstain. (C) WT or Glrx1−/− MTE cells were treated with 1 μg/ml LPS for 6 h, and NF-κB DNA binding was assessed by EMSA. NS, nonspecific binding. (D) WT or Glrx1−/− MTE cells were treated with 1 μg/ml LPS for 24 h, and concentrations of keratinocyte-derived chemokine (KC) (filled bars) and MIP-2 (open bars) were assessed by ELISA on culture media and corrected for protein content.