Ectopic expression of PKB regulates neutrophil differentiation ex vivo. CD34⁺ cells were retrovirally transduced with myrPKB, PKBcaax, or eGFP alone. (A) Retrovirally transduced CD34⁺ cells were cultured in presence of G-CSF to induce neutrophil differentiation. After 17 days of culture, transduced cells were separated from the nontransduced cells by FACS, and cytospins were made. Cytospins were stained with May-Grunwald Giemsa solution. (B) Data were expressed as the percentage of differentiated neutrophils. (C) Lactoferrin, (D) CD49d, and (E) CD14 expression was analyzed by FACS to determine neutrophil, eosinophil/monocyte, and monocyte development, respectively, in presence of G-CSF. (F) CD34⁺ cells were retrovirally transduced with myrPKB or eGFP alone. Retrovirally transduced CD34⁺ cells were cultured in absence of cytokines. After 17 days of culture, transduced cells were separated from the nontransduced cells by FACS, and cytospins were made. Cytospins were stained with May-Grunwald Giemsa solution. Data were expressed as the percentage of differentiated neutrophils. Error bars represent SEM.