Deposition of EP and CEP in normal and inflamed peritoneal tissues. Peritoneal tissues were isolated from mice at 72 hours after thioglycollate-induced inflammation (A, lower panels) or from nontreated mice as a control (A, upper panels). Immunofluorescent staining demonstrates EP or CEP (green fluorescence) and CD68 (red fluorescence). Magnifications ×200. (B) CEP and EP staining in noninflamed (green bars) and inflamed (blue bars) tissues were analyzed using Fiji software. (C) Neutrophil and macrophage accumulation in the peritoneal cavity during thioglycollate-induced peritoneal inflammation after anti-CEP mAb treatment. Mice were injected twice with anti-CEP mAb or IgM control (30 minutes before and 24 hours after injection of 1 mL of 4% thioglycollate). Neutrophils were isolated at 18 hours and macrophages were isolated at 72 hours after thioglycollate injection. Statistical analysis was performed using Student t test. (D) Immunoprecipitation with anti-CEP mAb. Mouse peritoneal exudate was isolated at 72 hours after injection of thioglycollate and incubated with anti-CEP antibody. Antibody-bound fraction was separated by Laemmli SDS gradient electrophoresis (4% -15%). Fg was detected by mass spectrometry in the major bands isolated by immunoprecipitation with anti-CEP mAb. (E) Western blot analysis with anti-CEP pAb after immunoprecipitation with anti-Fg mAb. The same peritoneal exudate was immunoprecipitated with anti-Fg polyclonal antibody and the isolated fraction was analyzed by western blot with anti-CEP polyclonal antibody. Fg, fibrinogen; IP, immunoprecipitation; mAb, monoclonal antibody; n/s, not significant; pAb, polyclonal antibody; WB, western blotting.