Figure 3

DNA-binding patterns of PITX3 wild-type and mutant proteins. EMSA (left panel) showed alterations in the binding patterns of S13N, G219fs and K111E proteins in comparison to wild-type. The normal pattern of WT PITX3-DNA binding consisted of several bands with the low mobility complexes likely representing homo- or hetero- dimer DNA interactions (black arrows) and the fast moving band corresponding to PITX3 monomers binding DNA (black arrowhead); all complexes disappeared after addition of anti-PITX3 antibody to reaction (right panel; supershift band is indicated with black asterisk). No DNA-protein complexes were observed for K111E mutant and some low mobility complexes (indicated by black arrows) were not present in EMSAs performed with S13N or G219fs mutant extracts. Western blot (bottom) confirms the presence of PITX3 forms in the corresponding cells.