To establish a persistently infected culture, 1610 7 ISE6 cells were plated in a 150 cm 2 flask and incubated for 24 hours at 32uC. Cells were then infected at an MOI of 10 with LGTV TP21. Cells were passaged every two to three weeks as needed and supernatants collected for viral titer via immunofocus assay to confirm productive viral infection. Samples for immunofluorescence and electron microscopy were prepared at multiple time points post-infection. No differences were noted between samples collected at various time points. Representative images for figures depicted were acquired between 90 and 260 days post infection.