Figure 5
From: Herpes simplex virus UL56 interacts with and regulates the Nedd4-family ubiquitin ligase Itch
siRNA knockdown of Itch has no apparent effect on the viral growth. Vero cells were mock-transfected (RNA [-]) or transfected with either negative control siRNAs (siCont) or siRNAs specific to Itch (siItch) for 48 h (A), and subsequently infected with indicated viruses (B, C). (A) siItch efficiently and specifically down-regulates protein levels of Itch, whereas levels of Nedd4 and β-actin were constant. (B) A multi-step growth curve in siRNA-treated cells (MOI 0.003 PFU/ml). Both wild-type (186) and ΔUL56Z viruses showed similar growth kinetics in siCont-treated cells and siItch-treated cells. (C) Expression of Itch and viral proteins with PY motifs (VP5, VP16, and UL56) by immunoblot. There was no difference in viral protein level between siCont- and siItch-treated cells. β-actin was used as a loading control.