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. 2013 Oct 21;288(48):34567–34574. doi: 10.1074/jbc.M113.512376

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© 2013 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 1. — NleH1 phosphorylates CRKL in vitro and co-immunoprecipitates with CRKL from mammalian cells. A, identification of CRKL (yellow spot) as a substrate of NleH1 kinase activity using a kinase substrate array (Invitrogen). B, recombinant NleH1, but not NleH1(K159A), phosphorylated recombinant CRKL in vitro. C, HEK293T cells were cotransfected with FLAG-CRKL and NleH1-HA. After 48 h, cell lysates were immunoprecipitated (IP) with either anti-HA or anti-FLAG antibody, followed by immunoblotting with anti-FLAG or anti-HA antibody. The protein expression levels of NleH1 and CRKL in cell lysates are indicated in the input fraction.