FIGURE 6.

TUG overexpression affects ubiquitylated protein abundance and p97 distribution. A, HeLa cells were transfected using the indicated amounts of a plasmid to express full-length, FLAG-tagged TUG. Cells were lysed 48 h after transfection, and immunoblots were done to detect the indicated proteins. An antibody to the TUG C terminus was used to detect endogenous and transfected TUG and to assess the level of overexpression. B, FLAG-tagged intact TUG (residues 1–550) or truncated TUG proteins (containing the indicated residues) were transfected HeLa cells. Cells were fixed, and immunofluorescence microscopy was performed to detect endogenous p97 and FLAG-tagged TUG, as indicated. Scale bar, 20 μm.