FIG 5.

Time course of RV infection in MA104 cells. Each panel shows a representative RV-infected cell fixed with 4% paraformaldehyde (PFA) at the indicated times postinfection. All samples were permeabilized and probed with MAb dNSP2_NT, MAb vNSP2_CT, and PAb NSP5 simultaneously using the appropriate secondary antibody. MAb dNSP2_NT was detected with goat anti-mouse labeled with Alexa Fluor 488 (green), PAb anti-NSP5 was detected with goat anti-guinea pig labeled with Alexa Fluor 633 (blue), and MAb vNSP2_CT was directly labeled with Alexa Fluor 568 (red); DAPI was used for Merge panels (light blue). (Right graphs) Fluorescence intensity graphs for colocalization analyses corresponding to the white arrows in the merged image for each time point with colors that correspond to those in each panel. (Bottom graph) Fluorescence intensity ratios of dNSP2 to NSP5 (white bars) and vNSP2 to NSP5 (black bars) in viroplasms (6 to 11 viroplasms per time point) were calculated at each time point.