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. 2005 Jul;79(13):8004–8013. doi: 10.1128/JVI.79.13.8004-8013.2005

FIG. 4.

FIG. 4.

Cleavage of NS4A/B. (A) Schematic representation of NS4A/B constructs. NS4A/B was derived by PCR and cloned in pCAGGS-HA. A schematic representation shows NS4B, NS4A/B, and NS4A/B in the presence of NS2B and NS3 (indicated by a triangle). (B) An immunoblot shows expression of these recombinant proteins in Vero cells. In the presence of NS2B and NS3, a complex pattern is obtained with bands coinciding with those obtained when only NS4B is transfected as indicated. Molecular mass markers (kDa) are at left. (C) Induction of ISRE-9-27-CAT after treatment with IFN. The percentage of CAT induction by IFN-β in the presence of the indicated expression plasmids is shown. CAT activities were determined as mean values from two independent experiments (P values of 0.03 to 0.05). (D) Activation of STAT1 by IFN in the presence of cleaved and uncleaved DEN proteins. GFP-tagged STAT1 protein was visualized by fluorescence microscopy in cells previously transfected with the indicated constructs and briefly stimulated with IFN-β. NS4A/B+2B+3, NS4A/B+NS2B+NS3. The presence of NS2B+NS3 is indicated by a triangle.