Fig. 4.

Akt can phosphorylate Foxa-2 on residue T156. Recombinant GST-Foxa-2, GST Foxa-2_T156A, or GST-crosstide [positive control (24)] were incubated in an in vitro kinase reaction together with precipitated HA-Akt or HA-Lck-Akt from HEK293 cell lysates and 0.5 μCi of radiolabeled [γ-³²P]ATP (Top and Middle). Untransfected HEK293 cell lysates served as control. Proteins were separated by SDS/PAGE, and phosphorylated proteins were visualized by autoradiography (Bottom). Equal loading was determined by Western blotting using an anti-Foxa-2, anti-GST, or anti-HA antibody.