Figure 5.

(A) Tight junction disturbance. Caco-2 cells grown on filter inserts were infected with Puumala virus at MOI of 0.1 and analyzed by confocal laser-scanning microscopy. At 48 h cells were fixed and stained for PUUV nucleocapsid protein (red) and zonula occludens protein 1 (green). Cell nuclei were stained by DAPI (blue). (B) Cytoskeleton rearrangements in infected Caco-2 monolayers. Caco-2 cells were infected with PUUV at MOI of 0.1. At different time points the cells were fixed with methanol-free formaldehyde and stained with antibodies against PUUV nucleocapsid protein (red) and actin (white). Cell nuclei were stained by DAPI (blue). (C) Leaks in Caco-2 cells monolayers at high MOI. Caco-2 cells grown on coverslips were infected with Puumala virus at MOI of 1.0. At different time points the cells were fixed and stained with antibodies against Puumala virus nucleocapsid protein (red) and zonula occludens protein 1 (green) and analyzed by confocal laser-scanning microscopy. Mock control is shown at 96 h.