Expression of MyrAKT restores erythroid differentiation in EML-D816V cells. (A) Phosphorylation of AKT in EML cells expressed different forms of AKT. EML-WT or EML-D816V cells transduced with vector, EGFP-AKT1, or MyrEGFP-AKT1 were serum starved for 3 hours before stimulation for 5 minutes with or without 250 ng/mL mSCF. Immunoblot was done with antiphospho-AKT (S473) antibody. Membranes were stripped and reprobed with anti-AKT antibody. Arrows represent the positions of hybrid (Myr) EGFP-AKT (top arrow), and endogenous AKT (bottom arrow). (B) Active form AKT promotes erythroid differentiation in EML-D816V cells. EML-WT cells and EML-D816V cells transduced with different forms of AKT were grown in Iscove modified Dulbecco medium 10 containing 10% BIT with Epo and/or mSCF for 12 days. Hemoglobin-positive cells were scored after benzidine staining. Data are shown as percentage of benzidine-positive cells compared with total cells in a field. Data are mean ± SD of 3 fields. A representative of 3 separate experiments is shown.
