Figure 2. In vitro generated LCs. (A)... | Rockefeller University Press

$Figure 2. In vitro generated LCs. (A) Wild-type (wt) BM cells were cultured for 3 d in the presence of GM-CSF and TGF-β. Dot plots represent the expression of DEC205 and EPCAM within the CD11c+MHCII+ compartment. Data are representative of three independent experiments. (B) Fractions I to IV as depicted in A were sorted and recultured for 24 h in the presence of GM-CSF and TGF-β. Plots show the expression of DEC205 and EPCAM for the indicated fraction and the proportion of cells in each quadrant. (C) Indicated fractions identified as in A were sorted by flow cytometry and morphology was assessed by microscopy. Data in B and C are representative of two independent experiments. Bars, 10 µM. (D) Quantitative real-time PCR for the expression of Cd207 in fractions I to IV sorted as in A. Values are normalized to Hprt and are the mean ± SD from three experiments. (E) Histograms represent CD103 expression on the indicated fractions identified as in A. Numbers represent the geometric mean fluorescence intensity (MFI) ± SD from three experiments. (F) Quantitative real-time PCR for the expression of Xcr1 and Batf3 in fractions I to IV sorted as in A. In vitro generated CD103+ DCs, and ex vivo isolated splenic CD8+ DCs were positive controls. Values are normalized to Hprt and are the mean ± SD from two experiments. (G) BM cells from reporter mice of the indicated genotype were cultured for 3 d in the presence of GM-CSF and TGF-β and expression of GFP was analyzed within the different gated fractions. Bar graphs show the MFI ± SD of the gated fractions from three mice per genotype. Data are representative of two independent experiments. (H) Quantitative real-time PCR for the expression of key transcription factors in fractions I to IV sorted as in A. Values are normalized to Hprt and are the mean ± SD from three experiments.$

Figure 2.

In vitro generated LCs. (A) Wild-type (wt) BM cells were cultured for 3 d in the presence of GM-CSF and TGF-β. Dot plots represent the expression of DEC205 and EPCAM within the CD11c⁺MHCII⁺ compartment. Data are representative of three independent experiments. (B) Fractions I to IV as depicted in A were sorted and recultured for 24 h in the presence of GM-CSF and TGF-β. Plots show the expression of DEC205 and EPCAM for the indicated fraction and the proportion of cells in each quadrant. (C) Indicated fractions identified as in A were sorted by flow cytometry and morphology was assessed by microscopy. Data in B and C are representative of two independent experiments. Bars, 10 µM. (D) Quantitative real-time PCR for the expression of Cd207 in fractions I to IV sorted as in A. Values are normalized to Hprt and are the mean ± SD from three experiments. (E) Histograms represent CD103 expression on the indicated fractions identified as in A. Numbers represent the geometric mean fluorescence intensity (MFI) ± SD from three experiments. (F) Quantitative real-time PCR for the expression of Xcr1 and Batf3 in fractions I to IV sorted as in A. In vitro generated CD103⁺ DCs, and ex vivo isolated splenic CD8⁺ DCs were positive controls. Values are normalized to Hprt and are the mean ± SD from two experiments. (G) BM cells from reporter mice of the indicated genotype were cultured for 3 d in the presence of GM-CSF and TGF-β and expression of GFP was analyzed within the different gated fractions. Bar graphs show the MFI ± SD of the gated fractions from three mice per genotype. Data are representative of two independent experiments. (H) Quantitative real-time PCR for the expression of key transcription factors in fractions I to IV sorted as in A. Values are normalized to Hprt and are the mean ± SD from three experiments.

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