Interaction in vitro; proteins produced as a recombinant fusion protein in bacterial system.

Only heterodimers of Scr and exd bind strongly to the DNA probe tested; alone, neither binds the DNA probe very well, if at all. Heterodimer formation is affected by mutations in the DNA target.

Scr and/or exd proteins were mixed with [32]P labeled fkh DNA. DNA-protein complexes observed by EMSA were used to deduce protein-protein interactions.

Interaction in vitro; proteins produced as a recombinant fusion protein in bacterial system.

Ternary complex of Scr and exd proteins with fkh DNA.

Interaction in vitro; proteins produced as a recombinant fusion proteins in bacterial system.

The exd-hth heterodimer and Scr bind cooperatively to DNA; neither exd-hth nor Scr bind the DNA probe tested on their own.

Scr was mixed with a preformed exd-hth heterodimer ("exd-HM") and [32]P labeled DNA containing a fkh egulatory element. DNA-protein complexes observed by EMSA were used to deduce protein-protein interactions.

Interaction in vitro; proteins produced as a recombinant fusion proteins in bacterial system.

exd and Scr bind DNA cooperatively as a heterodimer. exd alone does not bind the DNA, and Scr alone binds only weakly. Mutations to the binding sites disrupt the interaction.

Various combinations of exd and Scr were mixed with [32]P labeled DNA from a TfAP-2 regulatory region. DNA-protein complexes observed by EMSA were used to deduce protein-protein interactions.

Bait expressed in transfected HEK 293 T cells.

Prey expressed in transfected HEK 293 T cells.

The Scr E342G mutation, which abolishes Scr homodimerization, does not affect the Scr-exd interaction.

Interaction in vitro; proteins produced by in vitro transcription/translation.

exd, hth and Scr, in various combinations, were mixed with [32]P labeled DNA. DNA-protein complexes observed by EMSA, corresponding to Scr-exd heterodimers and Scr-exd-hth heterotrimers, were used to deduce protein-protein interactions.

Interaction in vitro; proteins produced as a recombinant fusion proteins in bacterial system.

Kd = 2.6mM

Interaction in vitro; bait and prey produced as recombinant fusion proteins in bacterial system.

Positive control; Gel free pull down assay using previously characterized biotin labeled DNA consensus sites. Complex was pulled down with streptavidin and Scr-exd heterodimer identified with an anti-FLAG antibody.