Characterization of TC-core puncta.

<p>A. Huh-7.5 cells were electroporated with TC-core RNA. At 72 hours post electroporation (hpe), cells were stained with FlAsH (1.25 µM, green) then fixed and stained for immunofluorescence using anti-core or anti-E2 antibodies (red). Scale bar is 10 microns. Fluorescence profiles of lined area in A. Distance is in pixels and intensity is in arbitrary fluorescence units (AFU). Percent colocalization of FlAsH stained puncta positive for anti-core or anti-E2 per cell is shown is upper left corner of fluorescence profile. B. Huh-7.5 cells were infected with HCV for 72 hours, treated with Click-iT in the presence of actinomycin D (1 µg/mL) to detect RNA, fixed, and probed with core antibody. C. <i>Left:</i> Single frame of an acquired fluorescent timelapse where DsRed exposures were taken every 2 seconds. <i>Right:</i> Single frame image where 5 particles were tracked and the trajectories are superimposed. Elapsed time between images is 100 seconds. <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002466#ppat.1002466.s016" target="_blank">Video S1</a>. Scale bar is 10 µm. D. TC-core RNAs were electroporated into Huh-7.5 cells, maintained for 72 hours, stained with ReAsh (red) and Ribogreen (1 µl/ml), and visualized by live cell fluorescence microscopy. A representative montage of TC-core puncta transporting with RNA is shown. Elapsed time is 1 minute and 50 seconds seconds and the distance traveled is approximately 8.32 µm. <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002466#ppat.1002466.s027" target="_blank">Video S12</a>.