Expression of EEN and MLL-EEN relocate EBP. (A-C) Subcellular localization of Myc-tagged EBP 1-767, EBP 1-358, and EBP 532-767. (D) EEN and EBP colocalize in the cytoplasm. (E) NLSEEN interacts with and recruits EBP from the cytoplasm into the nucleus. (F) NLSEENΔSH3 mutant unable to relocate EBP into the nucleus. (G) MLL-EEN partially relocates EBP into the nucleus. HeLa cells were transiently transfected with Myc-tagged EBP with FLAG-tagged EEN or MLL-EEN. EBP and EEN/MLL-EEN were visualized by anti-Myc and anti-FLAG antibodies, followed by FITC-conjugated antirabbit and rhodamine-conjugated antimouse antibodies, respectively. Nucleus in each case was visualized by DAPI staining. Overlay of DAPI-, FITC-, and rhodamine-stained images are shown in the last column. Original magnification, × 1000.