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. 2017 Oct 3;292(48):19890–19904. doi: 10.1074/jbc.M117.794271

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© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

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Figure 2. — ErbB4 ICD is deSUMOylated by SENP1, SENP2, and SENP5. A, COS-7 cells were transfected with HA-tagged ErbB4 ICD, His₆-tagged SUMO1, and FLAG-tagged wild-type or C603A mutant SENP1 as indicated. Cells were lysed in denaturing buffer, and lysates were incubated with Ni²⁺-NTA agarose to pull down His₆-SUMO conjugates. Whole cell extracts or pulldown samples were analyzed by Western blotting with anti-ErbB4. SENP1 expression was analyzed by Western blotting with anti-FLAG. B, COS-7 cells were transfected with HA-tagged ErbB4 ICD, His₆-SUMO1, or His₆-SUMO3 and FLAG-tagged SENP1 or SENP2 as indicated, and analyzed as in A. C, COS-7 cells were transfected as indicated with HA-tagged ErbB4 ICD, His₆-SUMO3, and FLAG-tagged SENP1, SENP2, SENP6, or SENP7 or GFP-tagged SENP3 or SENP5, and analyzed as in A. The effect of each SENP on deSUMOylation of ErbB4 ICD was analyzed in three to six independent experiments.