The E-cadherin/catenin complex is formed at the plasma membrane of alternatively activated macrophages. (A) BALB/c thio-PEMs and (B) human monocyte–derived macrophages were treated for 24 hours with the indicated stimuli, followed by total cell lysate preparation and immunoblotting with antibodies against E-cadherin or β-, α-, or p120-catenin. β-Actin was probed as a loading control. (C) Naive (N) and IL-4–steered BALB/c thio-PEMs and NMe ECs were lysed, and immunoprecipitation was performed using anti–E-cadherin or isotype control antibodies. Immunoprecipitates (left) and total cell lysates (right) were immunoblotted with antibodies against E-cadherin, β-, α-, and p120-catenin. (D) NMe cells and BALB/c thio-PEMs, after 24 hours of treatment with the indicated stimuli, were stained with anti–mouse E-cadherin or isotype control and analyzed by FACS. The naive and NMe histograms show an overlay of isotype staining (dotted) with anti–E-cadherin staining (bold). For comparison, all other histograms show an overlay of anti–E-cadherin staining in stimulated macrophages (bold) with anti–E-cadherin staining in naive macrophages (dotted). Isotype stainings were similar in all conditions. (E) BALB/c thio-PEMs, whether or not pretreated for 24 hours with DFMO, were IL-4–steered for an additional 24 hours, stained with anti–mouse E-cadherin ECCD2 or isotype control and analyzed by FACS. E-cadherin surface expression values (ΔMFI = [median fluorescence intensity]_{anti-E-cad staining} − [median fluorescence intensity]_{isotype staining}) of a representative experiment were plotted.