Iron overload phenotype in hepatocyte-specific Smad7-overexpressing mice. (A-B) Relative mRNA expression of hepcidin (Hamp) and several Tgf-β/Bmp target genes in the liver of Alb-Smad7-tg (n = 7) and control mice (n = 5), measured by quantitative real-time PCR. (C) Hematoxylin-and-eosin (H&E) staining (scale bar, 100 μm) and (D) Perls’ staining (scale bar, 20 μm) for iron depositions in liver of Alb-Smad7-tg and control mice. (E) Nonheme iron content in liver and spleen of Alb-Smad7-tg (n = 7) and control mice (n = 5). (F-G) Normalization of hepcidin and of phosphorylated Smad1 levels to liver iron in Alb-Smad7-tg and control mice. (H) Perls’ staining (scale bar, 50 μm) in the spleen of Alb-Smad7-tg and control mice. (I) Plasma iron levels in Alb-Smad7-tg mice (n = 7) compared with control mice (n = 3). (J) Hematological indices in Alb-Smad7-tg mice compared with control mice (n = 3 mice per group). Images are representative staining of 3 mice per group. Data were analyzed using GraphPad Prism software and results are shown as mean ± SEM. For the statistical analysis, a nonparametric distribution and the Mann-Whitney U test were used. *P < .05; **P < .005; ***P < .0005. Hb, hemoglobin; HCT, hematocrit; LIC, liver nonheme iron content; MCH, mean corpuscular hemoglobin; MCV, mean corpuscular volume; RBC, red blood cell.