FIG. 9.

Analyses of the interaction between phosphorylated AML1 and PEBP2β. COS-7 cells were transfected with pME18S (lane 1) or pME-FLAG-AML1 (lanes 2 and 3), together with pCMV-MK and pME-PEBP2β. The cells were treated for 5 min with EGF (lane 3) or left untreated (lanes 1 and 2) and harvested with lysis buffer. The cell lysates were precleared with protein G-Sepharose, mixed with anti-FLAG antibody, and rotated for 2 h at 4°C. Then, FLAG-AML1 was recovered on protein G-Sepharose beads. The washed beads were subjected to SDS-PAGE, followed by Western blotting with anti-PEBP2β antibody. Expression of each protein was confirmed by using 60 μg of total cell lysates. Arrows, phosphorylated and unphosphorylated AML1.