FIG. 6.

Analysis of N protein interactions in vivo by the mammalian two-hybrid assay. HeLa cells seeded on 35-mm-diameter dishes were cotransfected with 100 ng of the reporter plasmid p5xGal4SV40-luc (A) or pG5CAT (B), and 500 ng of pM or pM-N together with 500 ng of pVP16 or pVP16-N as indicated. Renilla or β-galactosidase reporter constructs were included in the transfection mixture to allow normalization of transfection efficiencies in the luciferase (A) and CAT (B) assays, respectively. The luciferase and CAT assays were carried out 48 h posttransfection. For the luciferase assays, data from three independent experiments are shown; the error bars indicate standard deviations. For the CAT assay, the arrow indicates the direction of migration by thin-layer chromatography (TLC).