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. 2006 Nov 28;400(Pt 3):573–582. doi: 10.1042/BJ20060831

Figure 6. An additional repression domain between amino acids 586 and 635.

Figure 6

(a) To confirm that the Gal4–TopBP1586–1435 fusion protein was a repressor, an experiment with a titration of plasmid concentrations was carried out and the results show the ability of this protein to activate transcription from the Gal4 DNA-binding site-containing promoter pG5luc in HEK-293T cells. The results are expressed relative to pG5luc values. (b) To determine whether this region was able to repress the TopBP1 transcriptional-activation domain, an additional Gal4 fusion plasmid was constructed as shown. (c) The ability of the encoded fusion protein to activate transcription from a Gal4 DNA-binding site containing promoter, pG5luc, was assayed in HEK-293T cells. The results are expressed relative to the Gal4 DBD-expressing plasmid pBIND. (d) The plasmids encoding the proposed fusion protein described in (b) were transfected into HEK-293T cells. Protein extracts were prepared from these cells and Western blots using an anti-Gal4 antibody were carried out. A representative blot is shown. The numbers to the left-hand side of the blot represent the marker proteins size in kDa.