FIG. 8.

Effect of Ezh2 on transcriptional activity of Gal4-Eed. Jurkat cells were transfected with a mixture of reporter and expression plasmids. (A) Plasmids used in the transfection experiments. The reporter plasmid was a luciferase gene pGL3-promoter vector containing an SV40 promoter with five Gal4-binding elements. The expression plasmids were as follows. The mammalian vector, pM1, was used for expression of either the Gal4 DNA-binding domain alone (Gal4) or a fusion of Gal4 DNA-binding domain with the wild-type Eed (Gal4-Eed). Plasmid pM1 expressing the N-terminal fragment of the human Ezh2 protein (N-Ezh2, aa 1 to 203) instead of Gal4 was also used. (B) Result of the transfection experiments. Expression plasmids (1 μg of Gal4 or Gal4-Eed and 3 μg of N-Ezh2) with 0.3 μg of luciferase reporter plasmid were used for cotransfections. The total amount of DNA, 4.3 μg, per transfection was adjusted with pM1. Two days after transfection, cells were analyzed for luciferase activity. The data shown represent the means ± standard errors calculated from three independent experiments.