Figure 5. Relative affinity of recombinant Zbed4 for the different oligonucleotides.

Zbed4 (20 µg) was incubated with 0.5 nM different 20-mer ssDNA (left panel) and ssRNA oligonucleotides (right panel) for 45 min and the whole reaction mixtures (20 µl) with the protein-DNA complexes were subjected to EMSA on 1% agarose gels run at room temperature in HEPES buffer, pH 8.2, at 20 mA. A. Agarose gel stained using SYBR Gold for detection of nucleic acid retardation. B. The same gel stained with Coomassie R-250 for detection of Zbed4 protein. Zbed4 (20 µg) and a single primer (0.5 nM) were applied separately to the gel as controls. 1 kb DNA ladder was used as a standard for nucleic acid size. As seen, Zbed4 binds only to DNA and RNA 20-mers that contain poly-Gs.