FIG. 7.

Activation of NF-κB by rHagB. (A) Phosphorylation of NF-κB. C57BL/6 peritoneal macrophages (3 × 10⁶ cells) were stimulated with rHagB (20 μg/ml) for 0 to 120 min. Equal amounts of whole-cell lysates were subjected to electrophoresis on a 10% Tris-HCl gel, and Western blots were performed using a specific Ab against the phosphorylated form of NF-κB p65 (Ser536) or NF-κB p105 (Ser933). Total NF-κB p65 in each sample was used as the equal loading control. Blots are representative of results obtained from three separate experiments. (B) Activation of NF-κB p65 translocation. C57BL/6 peritoneal macrophages (5 × 10⁵ cells) were stimulated with rHagB (20 μg/ml) for 0 to 120 min. Equal amounts of nuclear extracts were analyzed for NF-κB p65 activity using the TransAM assay. Data represent the mean ± standard deviation of three separate experiments.