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. 2014 Apr 7;211(4):669–683. doi: 10.1084/jem.20130977

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© 2014 Berres et al.

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Figure 7. — Expression of BRAF-V600E in early BM-resident DC progenitors in BRAFV600E^CD11c mice results in multi-systemic high-risk disease and is associated with high local cytokine expression and the recruitment of additional inflammatory cells. (A) Relative mRNA expression of BRAF-V600E in sorted BM lineage negative sca1⁺ c-kit⁺ (LSK), lin⁻ sca1⁻ c-kit⁺ CD34⁺ CD16/32⁺ granulocyte myeloid progenitors (GMPs), lin⁻ sca1⁻ CD135⁺ c-kit^high CD115⁺ monocyte and DC progenitors (MDPs), and lin⁻ sca1⁻ CD135⁺ c-kit^low CD115⁺ common DC progenitors (CDPs) and lung/liver CD45⁺ MHCII⁺ CD11c⁺ DCs in BRAFV600E^CD11c mice as assessed by mutation-specific qPCR (data normalized to total BRAF expression, no BRAF-V600E expression detected in corresponding cells of control mice, n = 3, pooled data of three independent isolations). (B and C) Relative numbers of viable singlet CD11b⁺ CD115⁺ Flt3⁺ pre-DCs and CD11c⁺ MHCII⁺ DCs among circulating viable, singlet CD45⁺ cells (B) and hemoglobin levels (C) in BRAFV600E^CD11c mice, BRAFV600E^langerin mice, and control mice (n = 4–10 per group, pooled data of four independent experiments, each data point represents one mouse). (D) Immunohistochemical analysis of liver and lung tissue sections stained with indicated mAb in BRAFV600E^CD11c mice (top) and control littermates (bottom; bars, 50 µm). (E) Absolute numbers of tissue-infiltrating total hematopoietic CD45⁺ cells, F4/80⁺ macrophages, total CD3⁺ NK1.1⁻ T cells, total CD3⁺ NK1.1⁻ CD4⁺ CD8⁻ T cells, total CD3⁺ NK1.1⁻ CD4⁺Foxp3⁺ T reg cells, total CD19⁺ B220⁺ B cells, total CD3⁻ NK1.1⁺ NK cells, and total SiglecF⁺ CD11c⁻ eosinophils in lung and liver of BRAFV600E^CD11c mice and control littermates as assessed by multicolor flow cytometry (n = 4–5 per group, representative of two independent experiments). (F) Local expression of indicated chemokines and cytokines in peripheral liver tissue was assessed by qPCR (data normalized to Gapdh expression, n = 4–5 per group). All data are shown as mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001.