Figure 4.

IC87114 attenuates the disturbances in protein disulfide isomerase (PDI) folding status and NOX4 expression. (a) Carbonylated proteins in the immunoprecipitates from an anti-PDI antibody on nonreducing gels. (b) High-molecular-weight complex formation was analyzed with an anti-PDI antibody. (c) Reduced and oxidized forms of PDI were analyzed as described in the Materials and methods. Endoplasmic reticulum (ER) lysates treated with 10 mM dithiothreitol (DTT) and 5 mM diamide for 15 min were used as standards for the reduced and oxidized forms of PDI, respectively. (d) Lung lysates were treated with or without 1 mM diamide for 15 min, and then after washout during the indicated time period, the reduced and oxidized forms of PDI were analyzed via immunoblotting. The mRNA (e) and protein expression levels of NOX4 (f) in lung tissues obtained from ovalbumin/lipopolysaccharide (OVA/LPS), saline (SAL) and OVA/LPS-treated mice administered 1 mg kg⁻¹ IC87114 and vehicle (0.05% dimethyl sulfoxide (DMSO) with saline). The lines with SAL and OVA/LPS represent two samples of the same condition. In (e) and (f), values are presented as the mean (±s.e.m.) fold changes relative to the control (SAL) group (n=6 animals per experimental group). Mice challenged with saline (SAL) were used as the control group. ^#P<0.05 versus SAL; *P<0.05 versus vehicle.