The present study investigated the effect of microtubules (MTs) and vimentin during dengue virus serotype 2 (DV2) infection. Immunostaining showed that DV2 infection induced MT and vimentin reorganization. Colocalization of DV2 antigens with MTs or vimentin were often observed in ECV304 cells. MT-disrupting agents could enhance DV2 release but did not affect other steps of virus replication. In contrast, disruption of vimentin inhibited DV2 infection. Our results suggest that an MT-dependent mechanism may not be necessary for DV2 infection, and MT disruption may promote DV2 release. However, vimentin is required for DV2 infection.