Figure 1.

Gating strategy and flow cytometric characterization of pulmonary macrophage populations. 1A: Lungs of C3H and C57 mice were dispase digested and enriched for CD45+ cells using MACS 3–26 wk following 12.5 Gy whole lung irradiation. Cells were stained with antibodies to CD45, Ly6G, CD11b, CD11c, F4/80, Ly6C and CD206 and analyzed by flow cytometry. A representative sample is shown. Pulmonary macrophage subpopulations were distinguished based on CD11b and CD11c expression. 1B: For each subpopulation, expression of F4/80, Ly6C and CD206 was characterized. Autofluorescence was also assessed in the CD45+, Ly6G− population and the expression of CD11b, CD11c, F4/80, Ly6C and CD206 determined in autofluorescent positive and negative subpopulations. Additionally, the CD45+, Ly6G+ population was analyzed for the expression of CD11b, F4/80 and Ly6C. 1C: Pattern of macrophage presence in lung following irradiation relative to known pathology and time points.