FIG 3.

Cellular localization of the VP1-412-mCherry mutants and EGFP fusion proteins. (A) For each mutant, the percentage of cells displaying the fusion protein localized in the nucleus (N > C), diffused (N = C), or localized in the cytoplasm (C>N) was scored for at least 300 cells in at least three independent transfections. No single mutation resulted in an alteration of protein localization. (B) Double mutants containing mutation of BR1 together with BR4 or BR5 resulted in a diffuse localization of the protein. Mutation of three (C) or more (D) BRs confirmed the importance of BR1, BR4, and BR5 for nuclear localization of the fusion protein. (E) Confirmation of NLS activity of BRs. Indicated BRs were cloned in fusion with EGFP and expressed by transfection of PT cells. Localization of the proteins was evaluated by confocal microscopy through EGFP fluorescence in comparison with DNA staining with Hoechst reagent. Images for each clone are representative of three independent transfection experiments.