Analysis of hepatic iron content and ferritin expression of mice with tissue-specific versus total IRP2 deficiency. (A) Relative total nonheme iron levels and (B) Western blot analysis of ferritin H- (indicated FTH1) and L- (indicated FTL) chain expression in the liver of Ireb2^Δ/Δ mice lacking IRP2 expression in all tissues (total) versus Ireb2^+/+ littermates, and in Ireb2^VilCre, Ireb2^AlfpCre, and Ireb2^LysMCre mice with selective IRP2 ablation in intestinal epithelial cells (IECs), hepatocytes (hepat.), or macrophages (macro.), respectively, versus their corresponding control littermates; the presence (+) or absence (−) of the Cre transgene is indicated. (A) Data are presented as average plus or minus SEM. Within each group, the iron level of control mice was set to 100%. Top histogram represents male mice; bottom, female mice. The number of animals per group (n) is indicated. P: Student t test. (B) A representative Western blot experiment is shown. The faint band below the ferritin H-chain signal likely corresponds to a truncated product. β-Actin was used as a loading control. (C) Hepatic levels of HAMP1 mRNA were determined by qPCR in Ireb2^Δ/Δ mice versus Ireb2^+/+ littermates, and in Ireb2^AlfpCre mice carrying (+) or not carrying (−) the Cre transgene. HAMP1 mRNA levels were standardized to β-actin mRNA expression. The number of mice per group (n) is indicated. Data are presented as mean plus or minus SD. Within each group, HAMP1 expression in control mice was set to 100%. In Ireb2^Δ/Δ mice, the antagonistic effects of liver iron loading (A) and microcytic anemia (Table 1) on Hamp1 expression may neutralize each other. However, a Student t test revealed no statistically significant change in HAMP1 mRNA levels in Ireb2^AlfpCre(+) mice with an iron-loaded liver (A) and without microcytosis (Table 1). The histogram shows data obtained from female mice; similar results were obtained from males.