Figure 3. VACV exhibits delayed replication kinetics and less dsRNA abundance when grown at 30°C compared to 37°C.

(A) BS-C-1 cells were infected (MOI=10) with VACV and then incubated at either 30°C or 37°C for the indicated times. For each condition, 150 randomly viewed cells were scored as either positive or negative for the presence of a virus factory of any size. The bars depict the average values and the error bars represent the standard deviations of three independent trials. (B) Using the same data from the previous figure panel, images of virus factories were acquired and then analyzed using ImageJ software. The total size of 25 randomly selected virus factories was determined for each condition. The bars depict the average values and the error bars represent the standard deviations. (C) Representative images of virus factories are shown for the 4 hr. and 8 hr. time points at both temperatures. Cell nuclei are labeled as “N” in the images, which are at 1,000× total magnification. (D) BS-C-1 cells were infected (MOI=10) with VACV and then incubated at either 30°C or 37°C for 4 or 8 hrs. Cells were stained for B5 protein at the cell surface (red) and cell nuclei/virus factories (DAPI; blue). Images were merged using ImageJ software, are at 400× total magnification, and representative of two independent trials. (E) BS-C-1 cells were infected (MOI=10) with VACV and then incubated at either 30°C or 37°C for 24 hrs. Cells were stained for dsRNA (red) and cell nuclei/virus factories (DAPI; blue). Images were merged using ImageJ software, are at 400× total magnification, and representative of three independent trials.