Adiponectin modulates inflammatory reactions via calreticulin receptor–dependent clearance of early apoptotic bodies
J. Clin. Invest. Yukihiro Takemura, et al. 117:375 doi:10.1172/JCI29709 [Go to this article.]

Figure 7
Adiponectin interacts with calreticulin on the macrophage cell surface. (A) Calreticulin is immunoprecipitated by histidine-tagged adiponectin (APN) from detergent-solubilized THP-1 membranes. Membrane fractions were incubated in the presence or absence of polyhistidine-APN and then precipitated with nickel resin. The resin was then treated with a molar excess of histidine to release precipitated proteins, and this material was subjected to SDS-PAGE followed by immunoblot analysis with anti-calreticulin (anti-CRT) and anti-adiponectin (anti-APN) antibodies. WB, Western blot. (B) Adiponectin prepared from E. coli (E-APN) was immunoprecipitated from detergent-solubilized THP-1 membranes by anti-calreticulin antibodies. THP-1 membrane fractions were incubated in the presence or absence of polyhistidine APN and then subjected to immunoprecipitation with anti-CRT or control IgG. Immunoprecipitated material was then subjected to SDS-PAGE, and Western blot analysis was performed with anti-APN or anti-CRT antibodies. (C) Adiponectin inhibits the binding of anti-calreticulin antibody to macrophages. THP-1 macrophages were preincubated with 200 μg/ml adiponectin (red) or vehicle (blue) for 60 minutes followed by incubation with anti-calreticulin antibody (10 μg/ml) for 60 minutes. Cells incubated with chicken IgY followed by treatment with FITC-conjugated secondary antibody served as control. Cells were then incubated with FITC-conjugated secondary antibody to anti-calreticulin antibody and analyzed by flow cytometry. *P < 0.05 versus vehicle (n = 3).