Fig. 6.

Binding specificity of Freud-2/DRE complexes. EMSA was done using purified recombinant GST-Freud-2 (GST-hF2), GST or no protein incubated with labelled 5-HT1A 3′ or 5′ DRE (A or B, respectively). A specific complex (lower arrowhead) was observed for GST-hF2 but not GST, and this complex was competed with the indicated molar excess of unlabelled primers (see Table I). The 3′ 19-bp and 5′ 16- and 17-bp primers effectively competed (at 100x), indicating that human Freud-2 specifically binds to sequences in common between these primers (Table I). To confirm the presence of Freud-2 in the complex, antiserum (2 μl) to Freud-2 C-terminal (Anti-F2) was added and a supershifted complex was observed in the presence of GST-hF2 (upper arrowhead).