Peritoneal macrophages from SHIP−/− mice produce IL-6. (A) IL-6 production of freshly isolated and purified macrophages from bone marrow, spleen, lung, and peritoneum of WT and SHIP−/− mice, analyzed by ELISA after a 6-hour culture. Data are mean ± SE of 3 identical experiments. (B) Subpopulations of peritoneal cells were identified by flow cytometry as macrophages (Mac-1+, F4/80+), monocytes (Mac-1+, F4/80−, Ly-6Chi), myeloid dendritic cells (CD11c+, Ly-6C+), and neutrophils (Mac-1+, Gr-1+, Ly-6C+) and analyzed for intracellular IL-6. Shown is the total number of each population that expressed intracellular IL-6. Data are mean ± SE of 3 animals. (C) IL-6 histogram of peritoneal macrophages (defined as Mac-1+, F4/80+) from WT (dashed line) or SHIP−/− (solid line) mice. Unstained cells are shown in gray. (D) Peritoneal macrophages identified with anti–Mac-1 and anti-F4/80 antibodies. (E) Total cells from 7 separate animals.
