FIG. 8.

Effect of ICP0 and its homologues on the SUMO-1 conjugation of PML. Using Lipofectamine PLUS, HEp-2 cells were (A) cotransfected with pPML(F) and either pUC9 (lane pUC), pp65-ICP0 (lane ICP0), or the pp65-homologue plasmids (remaining lanes) shown, and harvested into SDS-gel loading buffer at 24 h posttransfection and analyzed by Western blotting using MAb anti-F at a dilution of 1/5,000 or (B) cotransfected with pPML(F) and either pUC9 or pp65-ICP0 and treated with 5 μM MG132 at 24 h posttransfection and harvested over the hourly time course as indicated. The positions of unmodified and SUMO-1-conjugated PML(F) bands are indicated by the long and short bars, respectively, in panel A, and the arrow (A) indicates a background antibody-detected band present in untransfected cells.