PML Ib reduces accumulation of viral transcripts but not ICP0 half-life.

<p>(A) Determination of ICP0 and Us11 expression in HSV-1 infected PML stable cell lines was determined via RT-real time-PCR. Stable cell lines containing PML I, PML Ib, PML I+PML Ib, or vector alone were seeded at 5×10⁶ cells per well in 6-well clusters and exposed to 0.01 PFU/cell of HSV-1 strain F diluted in mixture-199 (Gibco/Invitrogen) supplemented with 1% calf serum. After 1 hr incubation at 37°C, monolayers were cultured at 37°C and cells were harvested for isolation of total RNA at the indicated times. Murine 18S ribosomal RNA was used as the endogenous control. Data are representative of 2 separate trials. (B) PML Ib expression does not enhance the degradation of ICP0. Cell expressing PML Ib or vector control were exposed to 0.1 PFU/cell of HSV-1 (F). 12 h post infection, cells were treated with cyclohemimide (CHX) and protein lysates were harvested at the indicated times to monitor the expression of ICP0 by immunoblot. β-Actin was used as a loading control.