Supershift of SRY1 and 2 sites with the SRY antibody.

Experiments were performed with biotin labeled probes in the same conditions as EMSA assays previously performed, blotted to nylon membranes and revealed with Streptavidin-HRP by chemiluminescence detection. Lines 1, 2 and 3 contain biotin-labeled WT SRY sites (1–3, respectively), with nuclear proteins from SRY transfected HEK293 cells and IgG mouse, and the same probes with nuclear proteins plus 0.4 μg of monoclonal SRY antibody (αSRY). Arrows indicate the DNA-protein complexes and the supershift in the SRY1 and SRY2 sites. The asterisks indicate non-specific bands, and the negative control of unspecific binding of the SRY antibody with biotin-labeled probes (without nuclear proteins) is shown in the three lanes at the right.