In vitro interaction between the AML1 mutants and PEBP2β/CBFβ.

The total cell lysates of COS-7 cells containing the same amount of FLAG-tagged wild-type AML1 (lane 1) or AML1 R139G (lane 2) were incubated with the cell lysates containing PEBP2β/CBFβ for 2 hours. These cell lysates were precleared by protein G–sepharose and incubated with the anti-FLAG antibody for 3 hours; this was followed by recovery of the FLAG-tagged protein on protein G–sepharose beads. The beads were subjected to SDS-PAGE and Western blotting with the anti-PEBP2β antibody. The arrow indicates migration of PEBP2β/CBFβ.