Loss of α-hemoglobin–stabilizing protein impairs erythropoiesis and exacerbates β-thalassemia
J. Clin. Invest. Yi Kong, et al. 114:1457 doi:10.1172/JCI21982 [Go to this article.]

Figure 5
Oxidative stress in AHSP^–/– erythrocytes. (A) Relative ROS levels in erythrocytes at baseline and with added H₂O₂. ROS were measured by incubation of cells with DCFH, which is converted by ROS to the fluorescent product 2′,7′-dichlorofluorescein (DCF). (B) Protein oxidation in erythrocyte lysates. Twenty micrograms of hemolysate was treated with DNPH for derivatization of carbonyl groups (by products of protein oxidation). Protein-associated DNP was detected by Western blotting. An identical blot probed with anti-actin (bottom) indicates equal protein loading in each lane. (C) Susceptibility to phenylhydrazine-induced hemolytic anemia. Drug was administered on days –1 and 0. Blood was first sampled on day 0, then hematocrit (top panel) and reticulocyte counts (bottom panel) were assessed daily until recovery. No abnormalities were detected in AHSP^+/– mice in the experiments described in panels A–C (not shown).