Discordant expression of Foxp3 in induced Tregs.

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Discordant expression of Foxp3 in induced Tregs. Discordant expression of Foxp3 in induced Tregs. (A) Naive CD4 T cells were isolated from Foxp3gfp mice and stimulated in vitro for 1.5 d in Treg-inducing conditions prior to microscopy. Cells were stained with DAPI for DNA, anti–β-tubulin Ab, and anti-GFP Ab. Conjoined sibling cells from in vitro Treg induction culture undergoing cytokinesis. Signal shown in the Foxp3 panel is both from GFP-Foxp3 emission and anti-GFP Ab. Six representative sibling pairs with asymmetric GFP-Foxp3 expression and two representative sibling pairs with symmetric GFP-Foxp3 expression are shown. Scale bar, 5 μm. (B) Left, Quantification of GFP-Foxp3 asymmetry in cytokinetic pairs with at least one GFP+ sibling (n = 30). Dotted line denotes asymmetry value of 0.2, where values ≥0.2 are considered asymmetric. Right, Pie chart summarizing frequency of cytokinetic pairs with asymmetric GFP-Foxp3 (33%). **p = 0.0017 compared with DNA. Bonnie Yen et al. ImmunoHorizons 2018;2:119-128 Copyright © 2018 The Authors