FIG. 6.

Restricted Rev100-Rev100 and Rev86-Rev86 interactions in vivo. (A) HeLa cells were transfected with pGAL-Rev and pRev-VP, pGAL-Rev100 and pRev100-VP, or pGAL-Rev86 and pRev86-VP for two-hybrid assay. The cells were harvested, and the CAT/β-Gal values were calculated. The CAT/β-Gal value obtained by transfection of pGAL-Rev and pRev-VP was arbitrarily set at 1. The actual amount of CAT produced by this basal transfection sample was more than 300 pg/ml. The results represent the means and standard deviations of three independent experiments. (B) Western blot analysis to determine the expression level of GAL4- and VP16-fused proteins. The mock sample (lane “mock”) received pSRα296 alone. As the first antibodies, mouse anti-GAL4 monoclonal antibody and rabbit anti-VP antibody were used, respectively. The band (∗) that migrates more slowly than the authentic Rev100-VP band may represent modified Rev100-VP protein.