Figure 4
From: Oxidative stress augments toll-like receptor 8 mediated neutrophilic responses in healthy subjects
Effect of H 2 O 2 on the R848-induced nuclear factor-kappa B (NF-kB) activation. Cells were treated with or without 50 μM H2O2, and then further treated with various concentrations of R848 for 60 min. Phosphorylated NF-kB p65 was assayed by a flow cytometer (A), and nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha (IkBα) protein levels were assayed by western blotting (B). Each band intensity was assessed by densitometry. Relative intensity was calculated as the ratio of the specific band intensity to that of each appropriate β-actin band intensity. (C) PMNs were treated with 1 μM R848 with or without 50 μM H2O2 in the presence or absence of MG-132, a proteosome inhibitor. After 24 hrs, the media were assayed for IL-8 by ELISA. All values were mean values ± SEM of three to five separate experiments, and analyzed by ANOVA followed by Bonferroni's test. **p < 0.01, compared with the values of control; ++p < 0.01, compared with the values of H2O2-pretreated and vehicle-treated group; NF-kB p65 = nuclear factor-kappa B p65; IkBα = nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha.