Fig. 4. Confirmation of predicted direct Dfd downstream genes by EMSA. (A-D) EMSA for four predicted direct Dfd downstream genes tested using no protein, translation lysate only (L) and lysate with Dfd protein (D). To test the specificity of binding of Dfd protein to the DNA fragments, competitor oligonucleotides for the individual Dfd-binding sites (DBS) were used either in their wild-type (wt) or mutant (mt) sequence versions. The black arrowhead indicates the specific DNA-protein complex containing Dfd protein. Asterisks indicate the unbound labeled probe. Predicted Dfd response enhancers of the following genes were used: Eip63E (A), frizzled 2 (fz2) (B), wg (C) and CG5756 (D).