Iron content and expression of iron-related genes in the liver. Four-month-old wild-type (+/+) and homozygous TfR2 Y245X mutant (m/m) female sibling mice, as well as 6-month-old wild-type, heterozygous (+/m), and homozygous TfR2 Y245X mutant male murine siblings, were treated with either PBS or iron-dextran for 3 weeks. (A) Iron content in the liver. Iron content in the livers was measured using bathophenanthroline sulfonate as described in “Materials and methods.” (B) Northern blot analysis showing expression of hepcidin in the liver. The intensity of each band was measured by an image analyzer, and the ratio of the intensities of hepcidin and β-actin was calculated. Setting the mean value of the hepcidin/β-actin ratio in PBS-treated wild-type and heterozygous mice as 1, individual values were calculated and shown in a bar graph. The mean values of each group are compared and shown in a bar graph on the right (mean ± SD, ^*P = .021). (C) Western blot analysis showing expression of TfR2 and ferritin in the liver. Both the antihuman TfR2 and the antihuman ferritin antibodies cross-reacted to their murine counterparts. The bands around 120 kDa seen in the top panels are nonspecific. To show the loading balance, protein staining of the membranes also is shown. The intensity of each band was measured by an image analyzer and shown in bar graphs.