EZH2β is localized to the nucleus and partners with SUZ12 and EED. (A) Subcellular localization of EZH2β. Antibodies against total EZH2, EZH2α or EZH2β were used to determine EZH2α or EZH2β localization by immunofluorescence of epithelial cells transduced with HIS/EZH2β or HIS/EZH2α. Labeling of the HIS-tag was performed to both confirm localization and control for expression. Nuclei are counterstained with Hoechst and cytoskeletal components labeled with phalloidin red. Images were taken at 100× magnification. White scale bar represents 10 μm. (B) EZH2β interaction with SUZ12 and EED. Immunoprecipitation from whole cell extracts harvested from epithelial cells transduced with HIS/EZH2α or HIS/EZH2β using an antibody against the HIS-tag were probed with antibodies against SUZ12 and EED. Five percent inputs of whole cell lysates were included as control of transduction and expression. These results demonstrate that EZH2β localizes to the nucleus and interact with PRC2 targets, which are two key features expected of a functional EZH2 isoform. EED: embryonic ectoderm development; HIS: histidine; EZH2: enhancer of zeste homologue 2; EV: empty vector; immunoprecipitation; SUZ12: suppressor of zeste 12.