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EWS requires > 1 RGG domain for efficient nuclear import. A panel of EWS constructs lacking multiple RGG domains was cloned into the pEGFP C3 vector (BD Bioscience Clontech) and transiently transfected into HeLa cells using GeneJuice (Novagen). Removal of >1 RGG domain prevents efficient nuclear entry of a GFP-tagged EWS construct.
