FIG 3.

Downregulation of p62 during HSV-1 infection is independent of viral replication but dependent on immediate early gene expression. (A) HEL cells were infected with HSV-1(F) or the ΔICP8 virus (3 PFU/cell), and whole-cell lysates were collected at 3, 9, and 24 h postinfection. Equal amounts of protein were loaded for immunoblot analysis using antibodies against OPTN, p62, ICP0, and ICP8. ICP0 served as a control of infection, whereas VP16 served as a control for viral-genome replication. β-Actin served as a loading control. (B) HEL cells were infected with HSV-1(F) (5 PFU/cell), the ΔICP27 mutant (5 or 50 PFU/cell), or the ΔICP4 mutant (5 or 50 PFU/cell), and whole-cell lysates were collected at 9 h postinfection. Equal amounts of protein were analyzed by immunoblot analysis using antibodies against p62 and ICP0. β-Actin served as a loading control. (C) HEL cells were infected with HSV-1(F) or the RF or ΔICP0 mutant at 5 or 10 PFU/cell, and whole-cell lysates were collected at 9 h postinfection. Equal amounts of protein were analyzed by immunoblot analysis, using antibodies against p62 and ICP0. β-Actin served as a loading control. All experiments were repeated three independent times, and representative Western blots are presented.