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. 2015 Jan 13;290(10):6495–6506. doi: 10.1074/jbc.M114.592717

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© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 9. — Direct binding of the Duox A-loop to the DuoxA N terminus. Purified GST-tagged DuoxA N terminus was mixed with synthesized biotin-labeled Duox A-loop in binding buffer. IP, immunoprecipitation. Then, streptavidin-coupled magnetic beads were added 2 h later. The material absorbed to the beads was eluted in Laemmli sample buffer. The eluents were subjected to SDS-PAGE followed by immunoblotting (WB) using a polyclonal antibody against GST. The strongest interaction was observed between Duox1(Aloop) and DuoxA1(N) relative to Duox2(Aloop) + DuoxA2(N), Duox1(Aloop) + DuoxA2(N), and Duox2(Aloop) + DuoxA1(N) (n ≥ 3).