Fig. 7.
AML-1/ETO disrupts nuclear matrix compartmentalization of PLZF and reduces its ability to bind to DNA.
(A) 200 ng vector, PLZF, AML-1/ETO, or PLZF plus AML-1/ETO was transfected in 293T cells. The cells were fractionated into cytoplasmic, nuclear, and nuclear matrix fractions. Immunoblotting with PLZF monoclonal antibodies was performed on each fraction and on the unfractionated whole cell lysates. (B) EMSA was performed on lysates from transfected 293T cells. Expression vectors in each transfection are indicated. The PLZF high-molecular–weight complex is indicated by the lower arrowhead, and the supershift generated by the addition of anti-PLZF monoclonal antibodies is indicated by the upper arrowhead. (C) Immunoblot of lysates from the cells used in the EMSA in B after normalization for protein concentration.