Figure 6

Disrupting vimentin inhibits virus release in BTV infected cells. A) Co-localization of untagged VP2 with vimentin in transfected cells. Top, untreated cells; Middle, cells treated with colchicine to disrupt microtubules; bottom, cells treated with acrylamide to disrupt vimentin. In each row VP2 and vimentin localisation are shown in green and red respectively. As expected, disruption of microtubules with colchicine causes a rearrangement in the localisation of vimentin. B) Effect of treating cells with colchicine on the release of virus from infected cells. Cells and culture medium were harvested at 4, 8 and 24 hours post infection with BTV-10 in untreated and colchicine (5 μg/ml) treated cells. Control samples are labelled 4c, 8c and 24c, respectively. The titre of cell associated and released virus for each timepoint normalised to 100% for untreated cells. Colchicine treatment resulted in a time dependent increase in the titre of cell associated virus and a time-dependent decrease in the titre of virus in the culture medium. D) As C but for cells treated with acrylamide (50 μM) to directly disrupt the vimentin network. Effect of acrylamide was similar but much faster and more dramatic than that observed by indirect disruption of vimentin through disruption of microtubules using colchicine. For C and D error bars indicate the standard error of three replicates of the experiments.