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. 2014 Jan 16;63(10):1550–1559. doi: 10.1136/gutjnl-2013-306253

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Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions

This is an Open Access article distributed in accordance with the terms of the Creative Commons Attribution (CC BY 3.0) license, which permits others to distribute, remix, adapt and build upon this work, for commercial use, provided the original work is properly cited. See: http://creativecommons.org/licenses/by/3.0/

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Migration of mouse Foxp3⁺ cells towards recombinant CCL20. Foxp3-GFP C57/BL6 mice were infected with Helicobacter pylori strain B128 7.13. After 3 weeks their splenocytes were harvested and pooled. CD4 enrichment was performed using EasySep separation and then GFP⁺ and GFP⁻ populations were sorted using flow cytometry. The CD4⁺FoxP3-depleted fraction and CD4⁺FoxP3-enriched fractions (A) were stained for CCR6 (B). Numbers at the top right denote the percentage of double positive events. Migration of these cells towards recombinant 500 ng/mL of recombinant CCL20 (rCCL20) was assessed using a 96-well Transwell plate (C). The migration index was calculated by dividing the number of migrated cells in the lower chamber of each well, by the average migration seen in the medium-only control wells. The graph shows means and SDs from three independent experiments each performed in triplicate. GFP, green fluorescent protein.