(A) Phosphorylation of AKT in human erythroid cells stimulated by EPO (10 U/mL). Day 8 cells were washed once in IMDM and lysed by adding an equal amount of a buffer containing 2% Triton X-100 after incubation for 1 hour in the presence of DMSO or various concentrations of LY294002. AKT was immunoprecipitated with specific AKT antisera. Immune complexes were resuspended in SDS-sample buffer and divided into two. Phosphorylation of AKT was detected using an anti-phospho-AKT antibody (A), and total AKT was detected using an anti-AKT polyclonal antibody (B). Bands were visualized by chemiluminescence.