FIG. 4.

EMSAs with different dephosphorylated SAF-box fragments. (A) One hundred seventy-five nanograms (0.05 pmol) of linearized SV40 DNA was incubated with 9, 28, and 46 pmol of dephosphorylated DEK fragments (lanes 2 to 4, 5 to 7, 8 to 10, 11 to 13, and 14 to 16, respectively) or without protein (input, lane 1) for 1 h at 37°C. Nucleoprotein complexes were analyzed on 0.6% agarose gels in 0.5× TBE (pH 7.8; see Materials and Methods) and visualized by ethidium bromide staining. The fragments used are indicated above the blots. (B) Map showing the identified DNA binding domain of DEK. Dark grey boxes, highly acidic regions; light grey box, SAF-box, a potential DNA binding domain; black box, nuclear localization sequence (NLS). wt, wild type.