Figure 7.

Synthetic double-DZFs interact in mammalian cells. (A) Interaction specificities of synthetic DZFs used to construct double-DZFs. Interaction specificities shown (arrows) were determined using the B2H system and the mammalian cell-based ‘activator reconstitution' assay as reporters (Supplementary Table 1 and Figure 1). Red X's indicate no significant interaction was detectable in either system. (B) Schematic of synthetic double-DZFs. Inter-DZF linkers are shown as black lines and are described in Materials and methods. (C) Interaction of double-DZF pairs assessed using the mammalian cell-based activator reconstitution assay. Controls testing the interaction of single DZF pairs and of an intact synthetic VEGF-A activator (consisting of the DBD that binds the VEGF-A gene covalently fused to the NF-κB p65 activation domain) are also shown. Fold-stimulation of VEGF-A protein expression was calculated using ELISA measurements of secreted VEGF-A that were performed at least three times. Bars shown represent mean fold-activations of VEGF-A expression and error bars indicate standard errors of the mean.