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. Author manuscript; available in PMC: 2014 Jan 1.

Published in final edited form as: Mucosal Immunol. 2012 Dec 12;6(4):847–856. doi: 10.1038/mi.2012.123

One to 2.5 million FACS-sorted pre-μDCs (CD45.2) were injected into CD45.1 recipients and analyzed on day 4 and day 6 or 7. Day 4: n=4 from two independent experiments. Day 6 or 7: n=8 from five independent experiments. * p<0.05, ** p<0.01, *** p<0.001 by Student’s t-test. a) CCR9, α4β7 and B220 expression on total pre-μDC-derived cells (CD45.2) in SI IEL (left) and LP (right) on day 4 after transfer. Numbers indicate percentage cells. Data shown are from one of four recipients with similar results. b) CD103 and CD11b on pre-μDC-derived CD11c⁺ cells and host CD11c⁺ cells on day 7. The plot shows data from one of eight recipients with similar results. c) Pre-μDC-derived CD11c⁺ cells (per million input cells) as percentage of host CD11c⁺ cells in different tissues on day 4 and day 6 or 7. Symbols of the same shape and color represent tissues from the same animal. d) Pre-μDC-derived CD103⁺CD11b⁻ and CD103⁺CD11b⁺ cDCs from SI LP or MLN and CD8α⁺CD11b⁻ and CD8α⁻CD11b⁺ cDCs from spleen are shown as percentage of the corresponding subset from the host on day 4 and day 6 or 7 after transfer. e) Aldefluor staining of pre-μDC-derived and host CD11c⁺ cDCs from MLN and spleen. Negative control staining in the presence of ALDH inhibitor DEAB is shown for comparison. Data are representative of three independent experiments with similar results. f) Pre-μDCs were sorted from Flt3L-treated mice and pre-incubated with 250 μg of either α4β7-blocking antibody (DATK32) or isotype control for 10 minutes at room temperature before transfer into congenic recipients. Recipient mice received 250 μg of α4β7-blocking antibody or isotype control every 12 hours and were sacrificed on day 4. Pre-μDC-derived total CD11c⁺ cells were calculated as percentage of host CD11c⁺ cells. Data show combined results from analyses of three mice from two independent experiments. Error bars show SEM. * p<0.05 by Student’s t-test.

One to 2.5 million FACS-sorted pre-μDCs (CD45.2) were injected into CD45.1 recipients and analyzed on day 4 and day 6 or 7. Day 4: n=4 from two independent experiments. Day 6 or 7: n=8 from five independent experiments. * p<0.05, ** p<0.01, *** p<0.001 by Student’s t-test. a) CCR9, α4β7 and B220 expression on total pre-μDC-derived cells (CD45.2) in SI IEL (left) and LP (right) on day 4 after transfer. Numbers indicate percentage cells. Data shown are from one of four recipients with similar results. b) CD103 and CD11b on pre-μDC-derived CD11c⁺ cells and host CD11c⁺ cells on day 7. The plot shows data from one of eight recipients with similar results. c) Pre-μDC-derived CD11c⁺ cells (per million input cells) as percentage of host CD11c⁺ cells in different tissues on day 4 and day 6 or 7. Symbols of the same shape and color represent tissues from the same animal. d) Pre-μDC-derived CD103⁺CD11b⁻ and CD103⁺CD11b⁺ cDCs from SI LP or MLN and CD8α⁺CD11b⁻ and CD8α⁻CD11b⁺ cDCs from spleen are shown as percentage of the corresponding subset from the host on day 4 and day 6 or 7 after transfer. e) Aldefluor staining of pre-μDC-derived and host CD11c⁺ cDCs from MLN and spleen. Negative control staining in the presence of ALDH inhibitor DEAB is shown for comparison. Data are representative of three independent experiments with similar results. f) Pre-μDCs were sorted from Flt3L-treated mice and pre-incubated with 250 μg of either α4β7-blocking antibody (DATK32) or isotype control for 10 minutes at room temperature before transfer into congenic recipients. Recipient mice received 250 μg of α4β7-blocking antibody or isotype control every 12 hours and were sacrificed on day 4. Pre-μDC-derived total CD11c⁺ cells were calculated as percentage of host CD11c⁺ cells. Data show combined results from analyses of three mice from two independent experiments. Error bars show SEM. * p<0.05 by Student’s t-test.