AML-1/ETO interacts with PLZF.

293T cells were transfected with 750 ng each of pCMV-AML-1/ETO and pCDNA-PLZF expression vectors. The cells were lysed and subjected to immunoprecipitations and immunoblotting as indicated. (A) Immunoblot with PLZF mouse monoclonal antibody. (B) Immunoblot with ETO C-terminal rabbit polyclonal antibody. (C) Immunoblot with AML-1 runt domain rabbit polyclonal antibody. The lanes of all 3 panels correspond to similar experiments. Lane 1, direct immunoblot of cell lysate. Lanes 2-6, extracts immunoprecipitated with mouse control antibody, rabbit control antibody, PLZF antibody, AML-1 antibody, and ETO antibody, respectively. (D) Immunofluorescence staining of 293T cells co-transfected with 100 ng PLZF and AML-1/ETO expression vectors. DAPI, nuclear staining. ETO, AML-1/ETO located using ETO polyclonal antibodies and Texas red–conjugated secondary antibodies. PLZF was detected using monoclonal antibodies and FITC-conjugated secondary antibodies. Ov, overlay of images of PLZF and AML-1/ETO. Magnification, 400×.