CysC–APN complex in serum and in vitro.
(A) Human serum was immunoprecipitated by the anti-APN antibody, followed by immunoblotting with either the anti-APN or anti-CysC antibody. Whole cell lysate (WCL) of HEK293T cells overexpressing both myc-tagged APN and CysC was used as a positive control. Experiments were repeated more than five times, and representative data are shown.
(B and C) Cell lysate expressing myc-APN and FLAG-CysC was used for the immunoprecipitation and immunoblotting. FLAG-CysC was co-immunoprecipitated with myc-APN by the anti-myc antibody (B), and myc-APN was coimmunoprecipitated with FLAG-CysC by the anti-FLAG antibody (C).
CON: control IgG, WCL: whole cell lysate, ip: immunoprecipitation, ib: immunoblotting. Experiments were repeated more than three times, and representative data are shown.