Increased hepcidin despite relative iron restriction in double-heterozygote mice. (A) Serum iron concentration (n = 4-6 mice per group) and (B) Prussian blue staining of liver samples (n = 3-4 mice per group) in WT, th3/+, and double-heterozygote mice. Nonheme liver iron (C), serum hepcidin (D), and hepcidin/liver iron (E) concentrations in 1-month-old mice (n = 3-4 per group), 5-month-old mice (n = 4-8 per group), and 15-month-old mice (n = 3-4 per group) in WT, th3/+, double-heterozygote mice. (F) Hepcidin mRNA in WT hepatocytes exposed in vitro to sera from WT, th3/+, and double-heterozygote mice (n = 5-7 mice per group). *P < .05; **P < .01 vs WT mice; †P < .05; ††P < .01 vs th3/+ mice. LIC, liver iron concentration.