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. 2010 Apr 21;285(25):19391–19401. doi: 10.1074/jbc.M109.084525

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 4. — PQBP1-Y65C shows a deficient interaction with WBP11. A, EGFP-WBP11 was transiently expressed in HEK293T cells with either HA-PQBP1 or HA-PQBP1-Y65C. Subsequently, the Lys, anti-HA immunoprecipitates (IP) and control immunoprecipitates (Ctr) were used for immunoblotting with anti-WBP11 antibodies (Ab). B, immunoblots show that PQBP1 but not PQBP1-Y65C co-immunoprecipitates with WBP11 from control- and GIH patient-derived lymphoblast cell lysates, respectively. C, PQBP1 forms a complex with Atrophin1 and WBP11. HEK293 cells were transiently co-transfected with PQBP1 and the indicated FLAG-tag plasmids. Cell lysates were immunoprecipitated with FLAG antibodies, resolved on SDS-PAGE, and immunoblotted with PQBP1 antibody (upper panel). Because WBP11 interacted strongly with PQBP1, only 10% of the total immunoprecipitates were applied on the gel for WBP11 samples (middle and lower panels show the expression of transfected proteins).