Effect of RAP-536 on iron overloading and Hamp expression in Hbb^th1/th1 mice.Hbb^th1/th1 mice (3-4 months old) were treated with RAP-536 or vehicle as in Figure 2, and age-matched wild-type mice treated with vehicle were used as additional controls. (A-C) Serum iron concentration, transferrin saturation, and serum ferritin concentration (n = 6-8 mice per group). (D-E) Tissue iron levels in liver and kidney (n = 5-7 mice per group). (F) Representative images of formalin-fixed sections from spleen, kidney, and liver stained with Perls’ Prussian blue reveal reduced iron content following RAP-536 treatment. Images were obtained with a 20× objective; bar = 100 µm. (G) Expression of Hamp and Bmp6 in liver determined by quantitative real-time polymerase chain reaction (n = 5-7 mice per group). Data are means ± SEM. ^#P < .05 and ^###P < .001 vs wild type; *P < .05 and **P < .01, RAP-536 vs vehicle.