Flow Cytometric Analysis of Mouse Mononuclear Cells

Mononuclear cells from the blood, bone marrow, and lung tissue were obtained, as previously described on p189 [20 (link),54 (link)], followed by flow cytometric analysis of the expression of surface markers of mouse mononuclear cells. Briefly, cell suspensions were stained with the following fluorophore-conjugated monoclonal antibodies: CD45 PerCP, CD31 APC, CD34 FITC, CD146 PerCP-Cy5.5, CD309 (Flk-1) APC, and CD117 (c-kit) PeCy7 (all Becton Dickinson, San Jose, CA, USA). Appropriate isotype controls were used. Labeled cells were thoroughly washed with PBS and analyzed on a FACSCanto II flow cytometer (Becton Dickinson, San Jose, CA, USA) using FACS Diva software. At least 100,000 events were recorded for each sample.