GSK360A-mediated mobilization of stored iron in the spleen is blocked in ISAM mice. (A) Berlin-blue staining detected iron deposition in spleen sections of the control or ISAM mice, 2 days after the final injection of GSK360A (Figure 4A). The scale bar indicates 50 μm. (B) The percentage of Berlin-blue–positive areas in each spleen section was quantified. (C) Serum concentrations of iron were measured 2 days after the final injection of GSK360A (Figure 4A). Data from C.E.R.A.–treated ISAM mice were used as positive controls for panels A-C. (D) The mRNA expression in the duodenums was analyzed in control or ISAM mice at 6 hours after injection of GSK360A (50 mg/kg) or roxadustat (200 mg/kg). In each graph, the average mRNA expression levels in the vehicle-treated control mice were normalized to 1. ∗P < .05 and ∗∗P < .01 determined by the Tukey-Kramer test.