Long term treatment with HJV.Fc reverses anemia in a rodent model of ACI by modulating the hepcidin-ferroportin axis and by mobilizing iron. ACI in female Lewis rats was induced by intraperitoneal administration of PG-APS and animals were followed up for 3 weeks. Then, ACI rats were treated with either HJV.Fc protein (20 mg/kg; ACI/HJV.Fc) or vehicle alone (ACI) by intravenous administration twice weekly over 28 days. (A) Hamp mRNA relative to Gusb mRNA expression in the liver, (B) hepatic Smad1 levels and Smad1/5/8 phosphorylation (pSmad1/5/8) as well as (C) Stat3 levels and Stat3 phosphorylation (pStat3), (D) serum iron levels and (E) the protein expression of ferroportin (FP-1) and ferritin in the spleen are shown after the termination of the experiment as detailed in the legend to Figure 3. (B-C) 1TBP18 was used as nuclear loading control and (E) β-actin as cytoplasmatic loading control. (F) Hemoglobin levels were determined in ACI rats once weekly starting with the initiation of HJV.Fc protein ♦ (light gray) or vehicle ● (dark) administration (day 0; 21 days after PG-APS injection). Results in panels A, B, C, D, F are reported as mean ± SEM, n = 10 for vehicle treated ACI rats and n = 10 for HJV.Fc treated ACI rats. Calculations for statistical differences between the various groups were carried out by Student t test. Exact P values are shown. (B,D) One representative Western blot is shown. Western blots used for densitometric quantification (B-C) are shown in supplemental Figure 3B.