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rHagB induces cytokine mRNA expression by murine macrophages. C57BL/6 peritoneal macrophages (3 × 106 cells) were stimulated with rHagB (20 μg/ml) for 0 to 24 h. Total RNA was isolated and the presence of mRNA for IL-10, IL-12p35, IL-12p40, IFN-γ, TNF-α, and β-actin in each sample was assessed by real-time PCR using SYBR green. β-actin was used as the endogenous control. The results are representative of three independent experiments.
