Fig. 4.

Inactivation of ROCK with Y-27632 inhibited DENV2-induced vimentin rearrangement and phosphorylation of vimentin at Ser71. ECV304 cells were pretreated with 10 μM Y-27632 or mock treated for 1 h at 37 °C prior to infection with DENV2. The cells were collected at different time points p.i. and then stained with the anti-vimentin antibody or analyzed by Western blotting with antibodies specific for Ser71-phosphorylated vimentin and vimentin, respectively. a Effect of ROCK on vimentin distribution. Immunostaining images show that Y-27632 prevented the rearrangement of vimentin aggregates into dense structures at the perinuclear area in DENV2-infected cells at 30 min, 1 h, 8 h, and 24 h p.i. Scale bars 10 μm. b Effect of ROCK on the phosphorylation of vimentin at Ser71. The results show that in Y-27632-treated cells, phosphorylation of vimentin at Ser71 was decreased at 30 min, 1 h, 8 h, and 24 h p.i. Each blot represents data from a minimum of six separate experiments