Figure 2. Gating strategy for the infected bone marrow-derived macrophages – upper panel, and the uninfected bone marrow-derived macrophages – lower panel.

After exclusion of doublets, the leukocytes are described as CD45 ⁺ . T-cells, B-cells, and NK cells are excluded (CD3–CD19–CD49b–), and macrophages are gated as Ly6G–CD11b ⁺ F4/80 ⁺ . Depending on the research question, there is the possibility to characterize pro-inflammatory macrophages as iNOS-expressing cells, and anti-inflammatory macrophages as ARG1–expressing cells. Furthermore, macrophages containing S. tm are characterized by expression of the red fluorescent protein (RFP) in the PE channel.