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. 2005 Jun 7;33(10):3303–3312. doi: 10.1093/nar/gki641

Figure 4.

Figure 4

Analysis of the interaction between APE1 and Kap α2 proteins: (A) 20 pmol of full-length APE1 (lane 1), ND33 (lane 2), ND20 (lane 3) or BSA (lane 4) were immobilized onto a nitrocellulose membrane, and probed with recombinant Kap α2 protein. Interaction was determined using Kap α2 antibody. (B) Far-western with full-length APE1 at 10 pmol (lane 1), 20 pmol (lane 2) or 40 pmol (lane 3); and ND20 APE1 at 10 pmol (lane 4), 20 pmol (lane 5), 40 pmol (lane 6) or BSA (lane 7) probed with Kap α2 protein followed by detection of Kap α2 with its antibody. (C) Far-western with full-length APE1 at 10 pmol (lane 1), 20 pmol (lane 2) or 40 pmol (lane 3); and ND33 at 10 pmol (lane 4), 20 pmol (lane 5), 40 pmol (lane 6) or BSA (lane 7) probed in the same way as in (B). (D) Interaction between APE1 and Kap α2 analyzed by His-tag pull-down assay. Kap α2 protein (15 μg), N-terminally tagged with histidine hexamer (His-tag Kap α2), was incubated with no protein (lane 1), 3 μg of WT APE1 (lane 2), ND33 APE1 (lane 3) or ND20 APE1 (lane 4) purified through Ni-NTA resin. The eluents were analyzed with anti-APE1 antibody. The arrows indicate the band positions of the full-length APE1 protein.

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