FIG 5.

Antiviral granules are formed around viral mRNA. HeLa cells were infected with WT-VACV at an MOI of 10. At 6 hpi, cells were treated with PBS, 500 μM arsenite (Ars), or 1 μM hippuristanol (Hipp) and fixed an hour later. Eighteen custom fluorescent oligonucleotides targeting the entire open reading frame of vaccinia virus F9L mRNA were hybridized prior to staining with antibodies to G3BP1 and with DAPI. Shown are example fields of view with individual channels for DAPI (blue), G3BP1 (red), and F9L mRNA (green) and merged images for PBS (i), Ars (ii), and Hipp (iii) treatments. N, cell nuclei; arrows, factories with AVGs; solid arrowheads, factories without granules; open arrowheads, SGs (bar = 5 μm). Charts display channel intensities of DAPI (blue), G3BP1 (red), and F9L mRNA (green) normalized for each channel between all images. The regions analyzed are marked with white bars on the merged images. The background level is indicated on each chart with a dashed line (0.2 RFU).