PLZF and FAZF interact in vivo. (A) Yeast two-hybrid assay. PJ69-4A yeast were transformed with a bait plasmid encoding full-length PLZF or the POZ domain of PLZF linked to the GAL4 DNA binding domain and the indicated prey plasmids containing full-length FAZF or PLZF or the POZ domain of PLZF linked to an acidic activation domain. Yeast colonies grown on leucine and tryptophan deficient media were expanded, and a liquid β-galactosidase assay was performed. Levels of β-galactosidase were normalized, setting the level of β-galactosidase obtained in the presence of a PLZF bait and PLZF prey plasmid to 1.0. The GAL4 protein acted as positive control for transcription. (B) In vivo–coimmunoprecipitation. 293T cells were transfected with PLZF or Flag-FAZF plasmids as indicated. Lysates from the cells were subjected to immunoprecipitation with the Flag M2 MoAb followed by immunoblotting with a PLZF polyclonal antibody.47