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. 2016 Nov 14;213(12):2553–2565. doi: 10.1084/jem.20160600

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© 2016 Wu et al.

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Figure 3. — Mafb-driven lineage tracing segregates macrophages from other populations. (A) Macrophages in MafB-mCherry-Cre × R26-stop-YFP × Zbtb46-GFP mouse BM, identified by surface markers as indicated (orange) after pregating as in Fig. S1 E, are compared with F4/80⁺ cells of similar immunophenotype (green) for expression of lineage-tracing marker YFP. Shown is one representative sample (n ≥ 4 animals over at least two independent experiments). (B) Alveolar macrophages in MafB-mCherry-Cre × R26-stop-YFP × Zbtb46-GFP mouse lung, identified by surface markers as indicated (red) after pregating as in Fig. S1 F, are compared with F4/80⁺ Siglec-F⁺ CD64⁺ cells of similar immunophenotype (blue) for expression of YFP. Shown is one representative sample (n ≥ 4 animals over at least two independent experiments). (C) MafB-mCherry-Cre × R26-stop-YFP × Zbtb46-GFP LPMs (purple) and SPMs (magenta), identified by surface markers as indicated after pregating as in Fig. S1 G, are compared for expression of Zbtb46-GFP and YFP in two-color histograms. Shown is one representative sample (n ≥ 3 animals over at least two independent experiments). (A–C) Numbers indicate percentage of cells within the indicated gate, and dotted gray lines show fluorescent signal measured in non-mCherry and non-YFP control samples. (D) Macrophages (identified as Zbtb46-GFP⁻ Ly-6G⁻ CD11b⁺ Mafb-mCherry⁺ YFP⁺, which were uniformly M-CSFR⁺), distinguished as LPM (F4/80^hi) or SPM (F4/80^lo), and DCs (identified as Zbtb46-GFP⁺ CD11c⁺) were sorted from the peritoneum of MafB-mCherry-Cre × R26-stop-YFP × Zbtb46-GFP mice and concentrated by Cytospin for morphological assessment by Wright–Giemsa staining. Shown are two representative cells for each population (n = 3 animals over two independent experiments). Bar, 20 µm. (E) DC-SIGN⁺ cells in the inguinal LNs of mice treated intravenously with vehicle (saline) or 10 µg LPS, identified by surface markers as indicated after pregating as in Fig. S1 H, are compared for expression of Zbtb46-GFP and YFP. Shown is one representative sample for each group (n ≥ 2 animals per group over two independent experiments). Numbers indicate percentage of cells within the indicated gate.