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. 2021 Mar 29;12:645741. doi: 10.3389/fimmu.2021.645741

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Copyright © 2021 Yang, Kwon, Kim, Im and Lee

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Microbial stimulation restores peripheral pool of Tγδ17 cells. Single cell suspension of indicated organs from SPF, GF and conventionalized GF B6 mice were analyzed by flow cytometry. (A) Graph shows statistical analysis of absolute number of thymic Tγδ17 cells at indicated ages. (B, C) Three and six week-old SPF and GF mice were analyzed using flow cytometry. (B) Pie charts show mean frequencies of each Vγ TCR among total Tγδ17 cells in indicated tissues from SPF and GF 3-week-old B6 mice (N = 3). (C) Graphs show statistical analysis of Vγ6⁺ and Vγ4⁺ Tγδ17 cells in indicated tissues from SPF and GF mice (N = 3). (D–G) GF mice were conventionalized by co-housing with SPF mice for 6 weeks (ConvGF) and analyzed. (D) Experimental scheme is shown. (E) Representative dot plots show Tγδ17 cells in the lung and SI-LP from SPF and ConvGF mice. Graph shows statistical analysis of absolute number of Tγδ17 cells indicated tissues. (F) Representative dot plots show thymic Tγδ2 cells and graph shows statistical analysis of their absolute numbers. (G) Representative dot plots show IEL Tγδ1 cells and graph shows statistical analysis of their absolute numbers. Numbers indicate frequencies of cells in adjacent gates and each dot represents an individual mouse. Error bars indicate ± SD. Pooled results from three independent experiments are shown. U.D, undetected. Unpaired two-tailed t-test was used. N.S, not significant; SPF, specific pathogen free; GF, germ-free; THY, Thymus; SPL, Spleen; PLN, peripheral lymph node; MLN, mesenteric lymph node; SI-LP, small intestinal lamina propria; IEL, intraepithelial lymphocytes.