N-terminal E4orf6 localization of diverse E4 mutant viruses.

(A) A549 cells were infected with the indicated mutant adenoviruses for 24 hours and proteins were resolved by immunoblot. RSA3 antibody shows various banding patterns for E4orf6/DBP, E4orf6, and E4orf6/7. * denotes E4orf6 frameshift product that is produced downstream of the common E4orf6 splice donor in dl1002. (B) A549 cells were infected with the viruses from A for 24 hours before immunofluorescence was performed. Cells were stained with antibodies against E4orf6 N terminal domain (RSA3, green) or cellular USP7 as a marker of viral replication centers (magenta). White scale bar denotes 10 μm. (TIF)