FIG. 7.

Effect of depletion of Sp100 isoforms containing SAND domains on IFN-β-mediated suppression of ICP0 expression. (A) Knockdown of Sp100 mRNA. Total RNA from HEp-2, and stable cell lines based on lentiviral infected HEp-2 cells, was extracted and subjected to semiquantitative RT-PCR for Sp100 isoforms and GAPDH. (B) Replicates of HEp-2 and stable cell lines expressing Sp100si-2 or NT2si were stimulated with 1,000 U/ml of IFN-β or left untreated as a control. Eighteen hours later cells were inoculated with 0.5 PFU of HSV-1 (strain 17+) per cell as described in Materials and Methods. After 3 h of infection, cells were harvested, and the nuclear fractions (30 μg/lane) were electrophoretically separated, transferred to nitrocellulose membranes, and probed with antibodies to ICP0 (top). Ezh2 was used as a loading control (bottom).