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. 2007 Jul 2;117(7):1933–1939. doi: 10.1172/JCI31342

Figure 5. Soluble HJV.Fc administration in mice decreases hepatic phosphorylated Smad1, Smad5, and Smad8 expression, decreases hepcidin expression, increases serum iron, increases liver iron content, and decreases spleen iron content.

Figure 5

129S6/SvEvTac mice received an i.p. injection of 25 mg/kg HJV.Fc or normal saline (control) 3 times weekly for 3 weeks. (A) Liver lysates were analyzed for phosphorylated Smad1, Smad5, and Smad8 (α–p-Smad1/5/8) expression by western blot. Blots were stripped and reprobed for expression of total Smad1 and β-actin, which were used as loading controls. Chemiluminescence was quantitated by IPLab Spectrum software for phosphorylated Smad1, Smad5, and Smad8 expression relative to total Smad1 expression. (B) Total mRNA was isolated from livers and analyzed by quantitative real-time PCR for hepcidin mRNA expression relative to GAPDH mRNA expression as an internal control. (C) Spleen membrane preparations were analyzed for ferroportin expression by western blot. Blots were stripped and reprobed for expression of β-actin, which was used as a loading control. (D and E) Measurement of serum iron (D) and transferrin saturation (Serum Tf sat; E). (F and G) Quantitation of liver (F) and spleen (G) tissue iron content. Results are expressed as mean ± SD, n = 3 mice per group; *P = 0.0497, †P = 0.003, ‡P = 0.01, ΧP = 0.004, ζP = 0.03, #P = 0.009 for HJV.Fc-treated mice compared with controls.