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. 2006 Aug;80(16):8019–8029. doi: 10.1128/JVI.02164-05

FIG. 4.

FIG. 4.

Effect of IFN-β treatment on expression of IE ICP0. (A and B) Replicate HEp-2, HEK293, and 293-S cell cultures were stimulated with 1,000 U/ml of IFN-β for 18 h or left untreated as a control. Half of each replicate was mock infected or 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 total cell extract and the nuclear fractions (30 μg/lane) were electrophoretically separated on denaturing polyacrylamide gels, transferred to nitrocellulose membranes, and probed with antibodies to ICP0 (A) or ICP4 (B). WB, Western blot. (C) HEp-2, HEK293, and 293-S cells were transiently cotransfected with ICP0 promoter-luciferase and stimulated 24 h later with 1,000 U/ml of IFN-β for 18 h or left untreated. Cells were harvested and fractionated on nuclear and cytoplasmic fractions. Firefly luciferase activity was measured in the cytoplasmic fractions and normalized to protein concentration. Responsiveness to IFN-β was calculated as the ratio of normalized luciferase activity of treated cells to that of untreated cells.