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. 2012 Apr 23;287(25):21012–21024. doi: 10.1074/jbc.M111.337865

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

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FIGURE 1. — PDZK1 specifically interacts with PLC-β3. A, strategies for identifying PLC-β-interacting PDZ proteins are shown. B, lysates of HEK293 cells transiently transfected with HA-PDZK1 were precipitated with GST fusion proteins containing the C-terminal heptapeptide of each PLC-β subtype (GST-β-tail). The precipitates were then subjected to Western blotting to determine whether the PDZ-binding motif of PLC-β interacts with PDZK1. C, HEK293 cells were transiently transfected with HA-PDZK1 and one of the PLC-β subtypes. PDZK1 was immunoprecipitated using the HA antibody, and the immunocomplexes were subjected to Western blotting to detect the co-immunoprecipitated PLC-β subtypes. D, HEK293 cell lysates were subjected to immunoprecipitation (IP) using the PDZK1 antibody, and the immunocomplexes were subjected to Western blotting to confirm the endogenous interaction between PDZK1 and PLC-β3.