Fig. 4.

Characterization of Hairy-interacting motifs in A. aegypti JH/Met-repressed genes. (A) EMSAs showing the binding of in vitro-translated Hairy with E-box–like motif “CACATG” and its flanks from the promoter region of the Met/Hairy-repressed gene AAEL000044. The specificity of the binding was confirmed by competition with unlabeled specific probe; the presence of Hairy in the DNA–protein complex was verified by a supershift with anti-AaHairy polyclonal antibody. (B) The binding capacity of in vitro-translated Hairy to variants of E-box–like motif was tested by mutating the CACATG core. Hairy binds to “CGCATG” and CGAATG, but not to “CAAATG” sites. (C) The motifs along with corresponding flanking sequences used for the EMSA assays.