Acrolein exposure induces protein-adduct formation and disruption of thiol status within the epithelium. ( A ) Lung sections were obtained 6 hrs after a single OVA challenge and acrolein exposure (day 14) and probed with anti-acrolein antibody (red) or DAPI (blue), and staining intensity was quantified using Metamorph software A control image without primary antibody (no 1º) is shown to demonstrate specificity. ( B ) Epithelial proteins collected by lysis lavage were labeled with the cysteine-reactive probe Iodoacetyl-LC-biotin, and biotin labeling was visualized by streptavidin blot and quantified using Image J software. ( C ) Reduced GSH was measured by HPLC in lung tissue homogenates. Results are expressed as mean ± SEM (n = 3-6/group) (*, p < 0.05).