FACS analysis of spleen, bone marrow, peripheral blood mononuclear cells (PBMCs), peritoneal macrophages (TEPMs) and bronchoalveolar lavage (BAL) cells of the 7.2
fms-EGFP TG mice. (A) In the spleen, EGFP is expressed mainly on adherent cells. Mechanically disaggregated spleen cells were grown overnight in TC dishes, and adherent and nonadherent cells were subjected to FACS analysis (top panels). Splenocytes were also stained with anti–F4/80 and anti–Mac-1 antibodies as described in “Materials and methods” (bottom panels). (B) Coexpression of EGFP and cell-surface markers examined by dual-color FACS analysis in bone marrow and PBMCs. For CSF-1R staining, cells were incubated with rat anti–mouse c-fms antibody, and stained with PE-conjugated goat anti–rat F(ab′)2. Other markers used were PE-conjugated anti–Mac-1 and anti–F4/80 antibodies. Quadrants were set based on the profiles from non-TG littermate controls. (C). Mac-1 and F4/80 staining of peritoneal macrophages and bronchoalveolar lavage cells. Cells were stained with anti–Mac-1 and anti–F4/80 antibodies. EGFP+ cells were gated and presented as black histograms. Dotted histograms represent isotype control staining. All data are representative of 3 separate analyses.
