Analysis of neutrophils from nonsplenectomized and splenectomized HbE/β-thalassemia subjects. (A) Representative morphological analysis of human neutrophils in the peripheral blood of HbE/β-thalassemia subjects. Analysis of neutrophil nuclei lobe numbers in (B) healthy (control) individuals, (C) nonsplenectomized HbE/β-thalassemia patients, and (D) splenectomized HbE/β-thalassemia patients. One hundred neutrophils from each individual were counted and classified into neutrophil subsets based on nuclear lobe numbers. (E) RT-qPCR analysis of PU.1 mRNA expression levels relative to housekeeping gene GAPDH in neutrophils isolated from the peripheral blood of healthy control individuals, nonsplenectomized (NS), and splenectomized (S) HbE/β-thalassemia (BE) patients. The statistical significance between the groups was evaluated with Student t test. Cells were visualized following Wright Giemsa staining (original magnification ×400).