Hepcidin-independent acute hypoferremia in mice. WT mice were injected with saline (ctrl), FSL1, or LPS (25 ng of ligand per g body weight) and analyzed 3 hours after injection. (A-B) Plasma iron levels and transferrin saturation were assessed. (C-F) Hepatic ferroportin, hepcidin, TNFα, and IL6 mRNA levels were determined by qRT-PCR and normalized to 36B4 mRNA levels. (G) The hepatic non-heme iron content was quantified as indicated. (H) Western blot analysis and quantification of ferroportin expression in the liver of injected 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. Results are representative of 3 independent experiments. *P < .05; **P < .01; ***P < .001; Student t test; n = 6 mice per group.