Additional file 1: of Activation and polarization of circulating monocytes in severe chronic obstructive pulmonary disease

Figure S1. Flow cytometry gating strategy to identify and characterize monocyte subpopulations. PMBCs were stained as described in the Methods section and at least 250,000 events per sample were collected. Singlets (red rectangle, panel a) were gated using the forward side-scatter area (FSC-A) vs height (FSC-H). From the singlets gate, monocytes (red oval, panel b) were gated using the FSC-A vs side-scatter area (SSC-A). The monocytes were further gated using CD14 vs CD16 and are indicated by the red boxes (panel c). The classical monocytes are CD14 + CD16-; the intermediate monocytes are CD14 + CD16+; and the non-classical monocytes are CD14DIMCD16+. From the classical gate, cells stained for CCR2, CCR5, CD163, CD206, and IL-13Ra1 are shown (panels i-m), and from the non-classical gate, the staining for CCR2, CCR5, CD163, CD206, and IL-13Ra1 are shown in panels d-h. The red histograms indicate the isotype control for each marker. The black histograms indicated the expression of each marker. (PDF 311 kb