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. 2016 Apr 27;291(25):13216–13228. doi: 10.1074/jbc.M116.721357

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© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 1. — Interaction between Top2 and Mus101. A, anti-FLAG-agarose beads were incubated with nuclear extract (NE) from Drosophila S2 cells overexpressing FLAG-Mus101. Endogenous Top2 and FLAG-Mus101 were detected in the immunoprecipitation product by rabbit anti-Top2 antibodies and rabbit anti-FLAG antibodies. MW, molecular weight. B and C, direct binding between Top2 and Mus101 was examined with purified proteins by pulldown assays. Mus101-His₆ was pulled down by HA-Top2 immobilized on anti-HA-agarose beads. In a reciprocal experiment using nickel-nitrilotriacetic acid (Ni-NTA) resin, HA-Top2 was pulled down by immobilized Mus101-His₆. Empty anti-HA-agarose beads and nickel-nitrilotriacetic acid resin were used as controls. HA-Top2 and Mus101-His₆ were detected with immunoblot by mouse anti-HA antibodies and rabbit anti-Mus101 antibodies, respectively.