FIG 3.

UbcH9 ubiquitin-conjugating enzyme does not play a role in the degradation of ICP0 at early times after infection. (A) Immunoblots of four HEp-2 cell clones selected after transduction of lentivirus encoding anti-UbcH9 shRNA, one clone transduced with a lentivirus expressing a nontarget shRNA, and wild-type HEp-2 cells. The electrophoretically separated proteins were reacted with antibody to UbcH9. β-Actin served as a loading control. (B) Schematic representation of the experimental design. (C and D) Stability of wild-type ICP0 (C) or RF mutant (D) in wild-type HEp-2 cells (lanes 3 to 5), nontarget cells (NT; lanes 6 to 8), or cells from clone 2 of panel A derived by transduction with shRNA against UbcH9 (lanes 9 to 11).