Fig 11.

Study of PML.I specific depletion. (A) Western blot analysis of HepaRG cells expressing EYFP-PML.I and control (shLuci) or PML.I-specific (shPML.I) shRNAs. (B) Western blot analysis of HepaRG cells expressing shLuci or shPML.I shRNAs using an antibody that detects all PML isoforms. (C) The relative plaque formation of wt and ICP0-null mutant HSV-1 in HepaRG cells expressing shLuci (control), shPML, or shPML.I shRNAs, as indicated. The data from several independent experiments were averaged and then plotted as means ± the standard deviations. Calculations were as described in Materials and Methods. (D and E) PML.I depletion delays the degradation of the other PML isoforms by ICP0. Control and shPML.I cells were infected with wt HSV-1 (MOI = 2) in absence or presence of MG132 or ICP0-null mutant HSV-1 (ΔICP0, MOI = 10) and harvested at hourly intervals until 7 h postinfection. Membranes were probed for PML, ICP4, ICP0, UL42, and tubulin.