Effect of ubiquitination on subcellular localization of Tax. Tax localization alone (A-F, top row) or together with DAPI to visualize the nucleus (A-F, bottom row) was examined by immunofluorescent microscopy in transfected 293 cells. Tax is expressed in nuclear foci in the absence of DNA damage (A) and is localized in the cytoplasm following UV irradiation (B). The addition of a UB tag at either the N- (C) or C- (E) termini caused Tax to be localized in the cytoplasm but treatment with LMB (D and F) sequestered UB-Tax and Tax-UB to nuclear foci. All images are shown at a magnification of 63×. The percentage of cells expressing nuclear foci (black bar) or cytoplasmic Tax (gray bar) was scored in five hundred transfected cells, in three independent experiments (G). Western blot showing expression of UB-Tax, Tax-UB, and native Tax proteins (H).