Figure 3

Lysosome inhibitor and proteasome inhibitor block the decrease of Itch. (A) Itch expression is stable in the absence of UL56. Vero cells were cultured for 24 h with or without cycloheximide (CHX, 100 μg/ml) and the lysates were analyzed for Itch expression. Cyclin D3 was used as a control. (B) Vero cells were mock-treated or treated for 12 h with either MG132 (10 μM) or chloroquine (CQ, 100 μM) and infected with wild-type HSV-2. Itch was detected in both cells treated with MG132 and those with CQ at 12 hpi, but only in those with CQ at 24 hpi. (C) The decrease of Itch is blocked by a lysosome inhibitor and partially by a proteasome inhibitor in cells stably expressing UL56. Vero, Vero-GFP-UL56, or Vero-GFP cells were mock-treated or treated with either MG132 (10 μM) or CQ (100 μM) for 24 h. β-actin was used as a loading control.