Hepcidin pretreatment blunts the LPS-mediated inflammatory response. TfR1 (A), IL6 (B), and TNFα (C) mRNA were analyzed by SybrGreen qRT-PCR in spleen macrophages isolated from ID mice pretreated with hepcidin (100 μg) or PBS before LPS injection (4 mice for each group). Gapdh was used as housekeeping gene to normalize gene expression. mRNA expression ratio was normalized to an ID (-hepcidin, -LPS) mean value of 1. (D) Serum IL6 levels were measured as described in “Serum IL6 levels” (4 mice for each group). Error bars indicate SE; *P < .05; and ***P < .0001.