Inflammatory hypoferremia and ferroportin protein downregulation are preserved in C326S ferroportin knock-in mice with a disrupted hepcidin/ferroportin regulatory circuitry. (A) Plasma iron level in C326S ferroportin mutant mice injected with FSL1 (100 ng/g body weight) for 3 hours. (B) Hepatic hepcidin mRNA levels were determined by qRT-PCR and normalized to 36B4 mRNA levels. Splenic (C-D) and hepatic (E-F) ferroportin mRNA and protein levels were analyzed by qRT-PCR and western blot in the same groups of mice. β-actin was used as loading control. Each lane in the western blot analysis represents the protein lysate obtained from a single mouse. Data are means ± SEM. *P < .05; Student t test; n = 6 mice per group.