FIG. 3.

UL47 shuttles between the nucleus and the cytoplasm in infected cells. (A) MDBK cells grown in a coverslip chamber were infected with jv46v at a multiplicity of 10. At 4 h, the cells were examined using a Deltavision RT imaging system and individual cells chosen for FLIP analysis. The laser module was then used to carry out sequential photobleaching events of the area denoted by the white oval. This area was exposed to the laser for 1 s every 10 s over a period of 7 min. An image of the field was acquired after each bleaching event to determine the loss of fluorescence in the nucleus of the cell. Arrows denote the control cell. (B) The relative fluorescences in the nuclei of the bleached and the unbleached cells in A were quantitated using NIH-image software and plotted against time. (C) MDBK cells infected with HSV-1 expressing GFP-NLS-VP22 in place of Wt VP22 were subjected to FLIP analysis as described for A.