Figure 4. BrdU multipulse/chase experiments showed viral DNA replication was arrested after irradiation.
Two plates of HFFs on glass coverslips were infected at an MOI of 5. One plate was irradiated at 75 J/m2 at 48 hpi and the other was left unirradiated Individual coverslips were pulse labeled with BrdU one h prior to harvesting (at −1,5,10 and 23 hpi, respectively). Coverslips were harvested at 0, 6, 11 and 24 hp irradiation. Cells were fixed and stained for BrdU incorporation (red) into viral RCs as described previously [68]. All images were captured using the same exposure time for direct comparison. Phase images of cells are pictured to show location of viral RCs within the nucleus (an example within each image is denoted by an arrow).