FIGURE 4.

Colocalization of VP2 with ORAOV1. (A) Hela or DF-1 cells were seeded on 24-well plates with coverslips and transfected with pDsRed-N1-vp2. Twenty four hours post transfection, cells were fixed and immunostained with rabbit anti-ORAOV1 polyclonal antibodies, followed by incubation with FITC-conjugated goat anti-Rabbit IgG antibodies. Nuclei were counterstained with DAPI (blue). The cell samples were observed under a laser confocal scanning microscope. (B) DsRed control is not colocalized with ORAOV1. Hela cells or DF-1 cells were transfected with pDsRed-N1. Twenty-four hours post transfection, cells were prepared and immunostained with rabbit anti-ORAOV1, followed by FITC-conjugated anti-rabbit IgG antibodies. (C) DF-1 cells were mock infected or infected with IBDV at an MOI of 10. Twelve hours after infection, IBDV pVP2/VP2 and endogenous ORAOV1 were probed with mouse anti-VP2 and rabbit anti-ORAOV1 antibodies, followed by FITC-conjugated goat anti-rabbit IgG and TRITC-conjugated goat anti-mouse IgG antibodies. The samples were observed under a laser confocal scanning microscope. (D,E) Localization of VP2 in the mitochondria. Both Hela and DF-1 cells were transfected with pDsRed-VP2 (D) or pDsRed-N1 (E) as controls. Twenty-four hours after transfection, cells were stained by MitoTracker Green for the mitochondrion. The cell samples were observed under a laser confocal scanning microscope.