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

COS-7 cells were transfected only with the pME/85 vector (lane 1). FLAG-tagged wild-type AML1 (lane 2), AML1 V105ter (lane 3), or AML1 R139G (lane 4) was coexpressed along with PEBP2β/CBFβ in COS-7 cells, and the cell lysates were precleared by protein G–sepharose and incubated with the anti-FLAG antibody for 3 hours. Then the FLAG-tagged proteins were recovered on protein G–sepharose beads. Washed beads were then subjected to SDS-PAGE and Western blotting with the anti-PEBP2β antibody.