Inhibition of sustained activation of Akt by wortmannin.

BAF3 cells expressing the D715 receptor were unstimulated or stimulated with G-CSF for 10 minutes prior to addition of wortmannin (100 nM) to the cultures. Whole-cell extracts were prepared at the indicated times and used for analysis of Akt phosphorylation (upper panel). The same membrane was incubated with anti-Akt antibody (lower panel).