FIG. 5.

Colocalization of HDAg-L and HA-NESI in the nucleus of transfected COS7 cells. COS7 cells were transfected with plasmid pcDNA3.1-HA-NESI encoding an HA-tagged NESI (A and B) or plasmid pECE-d-BE encoding the HDAg-L (C and D) or were cotransfected with both plasmids (E to H). Two days posttransfection, cells were fixed and subjected to indirect double immunofluorescence staining by using a mouse monoclonal antibody to the HA tag (A and G) and protein G-purified rabbit antiserum to HDAg (C and E) as primary antibodies. Panels B, D, and F are phase-contrast micrographs representing cells of the same fields as panels A, C, and E, respectively. HA-tagged NESI was visualized with Texas-red optics (A and G), and HDAg-L was visualized with fluorescein optics (C and E). A superimposed image of panels E and G is shown in panel H. Photographs were taken by epifluorescence microscopy.