FIG. 6.

Partial relocalization of HSV-1 AE into the cytoplasm is not sufficient to induce mRNA turnover. (A) 293T cells were transfected with plasmids expressing either WT HSV-1 AE or AE-NLSmut for 24 h and then subjected to IFA with anti-HA antibodies. DAPI staining was used to visualize nuclei. (B) 293T cells were transfected with a GFP reporter plasmid alone or together with the indicated AE construct for 24 h and then subjected to IFA with anti-HA (α-HA) antibodies. (C) 293T cells were transfected with increasing quantities of the indicated plasmids (100 to 300 ng). Total RNA was harvested 24 h posttransfection and Northern blotted with GFP and 18S probes. AE and muSOX levels were assessed by Western blotting with anti-HA antibodies. (D) The linearized pCDEF3 plasmid was incubated with aliquots of the indicated IVT protein for 1 or 15 min in degradation assay buffer at 37°C. The DNA was then extracted, resolved by agarose gel electrophoresis, and visualized by ethidium bromide staining. Vec, vector.