Figure 1. Analysis of tissues for the presence of monocytes.

For skin (A) and lung (C), doublet-cell excluded, live cells were plotted as CD11c vs CD11b or MerTK vs CD64. Live cells plotted as MerTK vs CD64 defined tissue macrophages as MerTK⁺CD64⁺. Gated MerTK⁺CD64⁺ macrophages were CD11c⁺ and CD11b⁺ (top panels). Continuing with the remaining MerTK⁻ CD64⁻ gate, DCs were identified as MerTK⁻CD64⁻CD11c⁺MHCII⁺ (second row panels), with both CD11b^hi and CD11b^lo subsets. Events captured neither in the macrophage or DC gate were then plotted to depict CD64 vs CD11b or CD11c vs CD11b, allowing us to identify monocytes and granulocytes (third row panels). Tissue monocytes were low SSC F480⁺Ly-6C⁺MHCII⁺ (fourth and fifth row panels). B) Blood monocytes were stained with anti-MerTK mAb, but lacked reactivity. Black line, anti-Mertk mAb; gray profile, Isotype control mAb. D) SSC overlay of tissue and LN monocytes, DCs and macrophages from gates shown in A, C. Data are representative of ≥ 3 experiments. Fig. S1 accompanies this figure.