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. 2011 Aug 1;208(8):1695–1705. doi: 10.1084/jem.20102657

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© 2011 Anandasabapathy et al.

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Figure 2. — Flt3L-responsive cells with dendritic morphology within meninges and choroid plexus. (A) Two-photon microscopy showing fluorescent cells in coronal brain sections of untreated CD11c-EYFP mice. EYFP⁺ cells were detected in choroid plexus (C.P.) and parenchyma (top left) and meninges and parenchyma (top right). Small panels at the bottom show the morphology of individual EYFP⁺ cells from choroid plexus, meninges (en face view), and parenchyma. (B) En face two-photon view of the brains of untreated CD11c-EYFP mouse brains. Blood vessels (red) were labeled by perfusion with DiI. EYFP⁺ (green) cells were detected in the meninges. Panels show a major blood vessel in the dura mater (top) and capillary blood vessels in the pia mater (bottom). (C) Two-photon microscopy coronal sections from the brain of untreated (top) and Flt3L-treated (bottom) CD11c-EYFP mice (green, EYFP). (D) Flow cytometric analysis of DCs in meningeal isolates. Dot plots show gated CD45^hiCD11c^hi DCs in CD45⁺ leukocytes of meningeal isolates and whole brain preparation in untreated or Flt3-treated B6 mice. Numbers indicate percentage of each cell type within total CD45⁺ cells. Bar graphs summarize the percentage of m/chDCs and microglia (MG) among CD45⁺ brain leukocytes in untreated versus Flt3L-treated mice in bulk brain and meninges (gating shown in Fig. S1 B). Bars show data from one representative experiment (n = 5 mice per group). Error bars represent the mean ± SD (n = 5). (E) Two-photon microscopy and flow cytometry of EGFP⁺ cells in the brain of I-A^b–EGFP transgenic mice. (microscopy) Observation in meninges 0–30 µm (left) and parenchyma 35–90 µm (middle) from the upper limit of the brain and in choroid plexus (right) from coronal sections of untreated I-A^b–EGFP mice (yellow, EGFP; red, collagen fiber; white, autofluorescence). Flow cytometry histograms show overlay of EGFP expression on microglia and m/chDCs, gated as in Fig. 1 B, from WT B6 (shaded area) and I-A^b–EGFP (line) mice. Data are representative of two independent experiments. Bars: (A, C, and E) 50 µm; (B) 100 µm.