FIG. 5.

Visualization of IEV by time-lapse confocal microscopy. At 1 h after infection of HeLa cells with 0.2 PFU of vB5R-GFP per cell, medium containing rifampin (0.1 μg/ml) was added to the cell monolayer. Approximately 12 h later, the rifampin was removed by washing with fresh medium. (A) Cells were viewed by confocal microscopy 2 h and 40 min later, and an image of one cell was collected every 10 s for 2 min. (B) After an additional 2 h, an image of the same cell was collected every 10 s. (C and D) HeLa cells were infected with 0.2 PFU of vB5R-GFP in the absence of rifampin, and imaging at 1 frame/6 s was started approximately 12 h later. In the upper left corner, the cumulative time elapsed (in seconds) after the start of image collection/video frame number is indicated. Arrowheads in each row point to the same IEV particle. In row B, the arrows point to virions extended from the cell on microvilli. In row D, the arrows point to tubules. N, nucleus. Scale bars, 50 μm. (The entire time-lapse videos are available at http://www.niaid.nih.gov/dir/labs/lvd/moss.htm.)