<p>Flow cytometric isolation of GFP<sup>+</sup> DP thymocytes from (<b>A</b>) Foxp3<sup>GFP</sup>, BAC-Foxp3<sup>Cre-GFP</sup> and (<b>B</b>) Foxp3<sup>IRES-GFP</sup> mice. Representative dot plots (from left to right) show presort analysis of CD4/CD8 expression among total thymocytes and CD25/GFP expression among CD25-enriched populations of gated DP cells, as well as postsort analysis of CD25/GFP and CD4/CD8 expression after flow cytometric isolation according to sort gates for CD25-enriched CD25<sup>+</sup>GFP<sup>+</sup> cells, as indicated. Lines with arrowheads illustrate the gating scheme. Numbers in dot plots indicate percentages of cells in the respective quadrant or gate. (<b>C</b>) Quantification of GFP<sup>+</sup> thymocytes. Percentages (left) and numbers (right) of GFP<sup>+</sup> DP cells (top) and GFP<sup>+</sup> CD4SP cells (bottom) from indicated Foxp3 reporter strains, revealed after postsort analysis as depicted in (<b>A,B</b>). All mice were six weeks old. (<b>D</b>) Numbers of GFP<sup>+</sup> DP thymocytes from eleven-week-old Foxp3<sup>GFP</sup> mice on the C57BL/6 (left) and BALB/c (middle) genetic background, as compared to age-matched BAC-Foxp3<sup>Cre-GFP</sup> mice (right). Dots and horizontal lines represent individual mice and mean values, respectively. * p < 0.05, *** p < 0.001, ns, non-significant (one-way ANOVA with Bonferroni’s multiple comparison post test).</p
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