Vero and ISE6 cells were seeded at 3610 4 cells/well in 8 well Labtek dishes (Nunc), infected with LGTV (MOI 10), and incubated for 48 hours at 37uC and 32uC, respectively. The cells were washed twice with PBS, fixed for 10 minutes in 4% paraformaldehyde (PFA)/5% sucrose in PBS, permeabilized with 0.1% Triton X-100/4% PFA for 10 minutes, washed with 50 mM glycine for 5 minutes, and blocked for 60 minutes with 2% bovine serum albumin (BSA)/PBS. Primary and secondary antibodies were incubated at a 1:1000 dilution in 2% BSA/PBS for 60 minutes each, with three five-minute washes with PBS between incubation steps. After washes, slides were dried and coverslips mounted using Prolong Gold Antifade with DAPI (49, 6-diamidino-2-phenylindole, Invitrogen). Confocal images were acquired using a Carl Zeiss LSM 710 confocal laser scanning microscope. TIFF files of individual channels and merge images were made using Bitplane Imaris 7.2 and images assembled for publication using Adobe Photoshop software.