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. 1999 Jun 8;96(12):6683–6688. doi: 10.1073/pnas.96.12.6683

Figure 2.

Figure 2

Binding of ZEB to CtBP mediates transcriptional repression. The region of ZEB corresponding to amino acids 1–1120, 302–903, and 700–776 were fused to the DNA binding domain of the yeast Gal4 protein and tested for its ability to repress transcription of the SV40 promoter/enhancer. Two micrograms of the Gal4 PM1 empty vector, 2 μg of wild-type G-ZEB-700–776 (amino acids 700–776), 2 μg of G-ZEB-700–776-mut (amino acids 700–776 with mutation of the CtBP site at 734), 2 μg of G-ZEB3 mut (amino acids 700–776 with mutation of CtBP sites at 705, 734, and 767), 3 μg of G-ZEB-302–903 (amino acids 302–903), 4 μg of G-ZEB-1–1120 (full-length ZEB), 2 μg of wild-type G-zfh-1–765-821 (amino acids 765–821) and 2 μg of G-zfh-1–765-821-mut (amino acids 765–821 with a mutation of the CtBP site at 792) were cotransfected with 0.8 μg of a reporter containing the SV40 enhancer/promoter (18). Transfection and assessment of CAT activity was performed as described in Materials and Methods.