Figure 8. Particle production in the presence of vaccinia virus inhibitors.

Hela cells were infected at a m.o.i. of 5-H-SFR and after adsorption, medium was replaced with fresh normal medium (brown rhombs) or medium containing 100 µg/ml AraC (red squares), 10 µg/ml IMCBH (black triangles) or 100 µg/ml Rifampicin (blue squares). Samples of the culture medium were collected every 12 hours, clarified, filtered and stored at 4°C until last point was collected. To determine the titer of SFPs, fresh BHK cells were infected with serial dilutions of the filtered supernatant, and GFP-positive cells were counted 24 hours later in the fluorescence microscope. Lower panels: Vaccinia virus Extracellular virus (EV) or cell associated (IV) production. Hela cell monolayers were infected at a multiplicity of 5 pfu per cell with W-H-SFR recombinant virus, and after 1 hour medium was replaced with medium containing AraC (100 µg/ml), rifampicin (100 µg/ml) or IMCBH (10 µg/ml). Virus was harvested at 24 hpi and Vaccinia virus titers in the culture medium (EV) (left panel) and cell lysates (IV) (right panel) were obtained by plaquing in BSC-1 cell monolayers and are represented relative to the control with no inhibitor.