FIG. 9.

MRV core particles colocalize with SGs in a manner dependent on viral gene expression. (A) HeLa cells were infected with MRV T1L ISVPs (1,000 PFU/cell). At 2 h p.i., cells were fixed and stained with rabbit anti-MRV core antiserum (left) and goat anti-TIAR antibodies (middle), followed by Alexa 594-conjugated donkey anti-rabbit IgG and Alexa 488-conjugated donkey anti-goat IgG. A merged image is shown (right). Confocal images were taken at 0.1-μm-slice intervals using a Leica SP5 X confocal microscope. The boxed regions in each image were amplified and are shown in insets in the merged image. (B) HeLa cells were infected with UV-inactivated T3D^C virions (1,000 PFU/cell). At 4 h p.i., cells were fixed and stained with rabbit anti-MRV core antiserum (left) and goat anti-TIAR antibodies (middle). A merged image is shown (right). The boxed region in the merged image was amplified and is shown in the inset. (C) HeLa cells were pretreated with 0.1 mg/ml puromycin for 1 h, incubated with T3D^C virions (1,000 PFU/cell) for 1 h, and then retreated with puromycin for an additional 10 h, at which point cells were fixed and stained with rabbit anti-MRV core antiserum (left) and goat anti-TIAR polyclonal antibodies (middle). A merged image is shown (right). The boxed region in the merged image was amplified and is shown in the inset.