In Hbbth3/+ mice, treatment with recombinant PEP1-Prx2 fusion protein ameliorates β-thalassemic anemia, beneficially affecting circulating red cells and erythropoiesis. (A) Hematological parameters: Hct, Hb, MCV, MCH, and retics in Hbbth3/+ mice with and without PP1Prx2 treatment (see Materials and Methods). Data are presented as means±SD (n=6). *p<0.05 compared with vehicle-treated Hbbth3/+. (B)
Upper panel. TAU gel electrophoresis of red cell membrane from WT and Hbbth3/+ mice with and without PEP1 Prx2 treatment. On the right, quantification of gel bands (OD) expressed as α-globin to β-globin ratio to Hb. Data are shown as means±SD (n=3); *p<0.05 compared with vehicle-treated Hbbth3/+ mice. Lower panel. Total and soluble Hb measured in hemolysates from Hbbth3/+ mice with and without PEP1 Prx2 treatment. Data are shown as means±SD (n=3); *p<0.05 compared with vehicle-treated Hbbth3/+ mice. (C) Iron staining (Pearl's Prussian blue) in spleen and liver from Hbbth3/+ mice with and without PP1Prx2 treatment. One representative image from the other six with similar results is presented. Lower panel. Quantification of iron staining on Pearl's Prussian blue spleen and liver from Hbbth3/+ mice with and without PP1Prx2 treatment mice. Data are shown as means±SD (n=6). *p<0.05 compared with vehicle-treated Hbbth3/+. (D) Cyto-fluorimetric analysis of maturation pattern of erythroid precursors from the bone marrow of Hbbth3/+ mice with and without PP1Prx2 treatment using the following surface markers: CD44 and TER119. Population I corresponds to pro-erythroblasts; population II corresponds to basophilic erythroblasts; population III corresponds to polychromatic erythroblasts; and population IV corresponds to orthochromatic erythroblasts. Data are presented as means±SD (n=6); *p<0.05 compared with vehicle-treated Hbbth3/+mice. (E) ROS levels in erythroid precursors from bone marrow of Hbbth3/+ mice with and without PP1Prx2 treatment. Data are presented as means±SD (n=6); *p<0.05 compared with vehicle-treated Hbbth3/+mice. (F) Annexin V+-7-AAD− sorted orthochromatic erythroblasts (population IV) from bone marrow of Hbbth3/+ mice with and without PP1Prx2 treatment. Data are presented as means±SD (n=6); *p<0.05 compared with vehicle-treated Hbbth3/+mice. Hb, hemoglobin; Hct, hematocrit; MCH, mean corpuscular hemoglobin; MCV, mean corpuscular volume. To see this illustration in color, the reader is referred to the web version of this article at www.liebertpub.com/ars