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. 2012 Oct 30;287(52):43741–43752. doi: 10.1074/jbc.M112.372318

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© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 2. — Usp9x directly interacts with SMN. A, immunoblotting (IB) of either FLAG or HA to examine overexpressed HA-Usp9x, FLAG-SMN, Gemins 2–8, or Unrip in 293T cell lysates. FLAG-SMN, Gemins 2–8, or Unrip was immunoprecipitated with the anti-FLAG M2 antibody. Precipitates were immunoblotted with an anti-FLAG or anti-HA antibody. Lane 1, cells overexpressing HA-Usp9x; lane 2, cells overexpressing HA-Usp9x plus a FLAG-tagged one of the proteins of the SMN complex as labeled on the far right. B, Coomassie staining of 1 μg of recombinant GST-Usp9x resolved by SDS-PAGE. C, Coomassie staining of 2 μg of recombinant His₆-SMN resolved by SDS-PAGE. D, immunoblotting of SMN (upper blot) and GST (lower blot) contained in GST (control) or GST-Usp9x pulldown assay.