Figure 2. Fluorescent labeling of recoated TLP.

A. Images of particles, adsorbed to coverslip, at each of three wavelengths and overlay. B. Analysis of recoated, purified TLPs by 12% SDS-PAGE. C. Fluorescent focus assay measuring effect of labeling on reacoated particle infectivity, with labeling as shown. D. Images of doubly labeled (VP7, green; DLP, red) recoated particles, 10 min and 45 min following addition to BSC-1 cells. Uncoated particles (red) are evident at the later time point. E. Time lapse sequence of triply labeled particles uncoating in BSC-1 cells. Time frames at 5 sec intervals from an arbitrary time point following addition of particles to cell. Pseudo-colors: VP7, blue; VP4, green; DLP, red. F. Graphic representation of time lapse sequence in panel E; images recorded at 1 sec intervals. Intensities (colors as in E) evaluated in a box moving with the DLP; orange curve: velocity of DLP.