Fig. 7.

Mutations acquired during mouse-adaptation do not influence RAVV VP40 budding in mouse cells. (A) Hepa1.6 cells were transfected with the indicated plasmids encoding FLAG-tagged wild-type or mutant RAVV VP40s. At 8 h posttransfection, 1,000 U of IFN-α/β/ml was added with fresh medium to the cells. After 40 h, the supernatants were harvested, and VLPs were purified through a sucrose cushion. Cells were lysed and examined together with the VLPs for protein expression levels (WCE, whole-cell extract). Anti-FLAG, anti-GFP, and anti-β-tubulin antibodies were used in both the WCE and the VLP blot. (B) Bands from panel A were quantified by using NIH ImageJ, and the ratio of VLP intensity to the WCE intensity was graphed after subtracting the value obtained for GFP.