Table 1.

Two-hybrid interaction between Eed and Ezh2 in Jurkat and COS cellsa

Bait constructLuciferase activity of VP16AD fusion construct in:
Jurkat cellsCOS cells
VP16VP16-Ezh2 (aa 39 to 159)VP16VP16-Ezh2 (aa 39 to 159)
Gal41.0 1.2 ± 0.21.01.5 ± 0.3
Eedb 0.8 1.3 0.70.4
Gal4-Eed0.3 ± 0.1147.0 ± 290.6 ± 0.1313.0 ± 12
Gal4-T1031A1.2b 1.4b 0.7 ± 0.10.4 ± 0.2
Gal4-T1040C1.0 ± 0.21.2 ± 0.30.7 ± 0.20.5 ± 0.2
  • a Cells were transfected with 0.5 μg of the reporter plasmid, bearing the firefly luciferase gene driven by the SV40 enhancer and minimal promoter containing five Gal4-binding elements, and two hybrid constructs (2 μg of each): the Gal4 DNA-binding domain alone (Gal4) or Gal4 fused to the wild-type or mutant Eed (Gal4-Eed, Gal4-T1031A, Gal4-T1040C), or nonfused Eed protein (Eed), and the VP16 activation domain either alone (VP161) or fused to N-terminal fragment of Ezh2 [VP16-Ezh2 (aa 39 to 159)]. Two days after transfection, the cells were analyzed for luciferase activity. Data represent the means ± standard errors calculated from three independent experiments.

  • b The experiment was repeated once.