Figure 6

Effect of Arf6/Q67L induced PI(4,5)P ₂ -enriched vesicles on virus production. (A) Western Blot analysis of HeLa (right) and BSR (left) cells transfected with HA-tagged Arf6/Q67L and infected with BTV-1 were analyzed for expression of viral (VP5 and NS2) and cellular (actin and tubulin) proteins 4 and 12hrs post-infection (lane,1). Controls consist of HeLa cells that have not been transfected but infected with BTV-1 (lane,2) and cells that have neither been transfected nor infected (lane, 3). The proteins were detected by specific antibodies. The western blots were repeated 2 times on three independent experiments. (B &C) Analysis of relative (B) and total (C) virus titres in infected HeLa (right) and BSR (left) cells, in the presence (+) or absence (−) of Arf6 mutant Arf6/Q67L. Cells infected with BTV-1 12 hrs post transfection were harvested at 4 and 12 hrs post-infection. For the relative virus tires, the total titre for each time was normalized to 100% for untreated cells. Bars indicate the standard error of three replicates of the experiments.