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. 2015 Oct 7;2015:352934. doi: 10.1155/2015/352934

Figure 4.

Figure 4

Th subset differences in STAT5 phosphorylation responses and STAT1 protein expression. (a) Colocalization of pSTAT5 within the 7-AAD stained nucleus after IL-2 stimulation in CD25+ T lymphocytes quantified by imaging flow cytometry as described in Section 2. Left: representative histograms of 7-AAD/pSTAT5. Similarity scores correlating 7-AAD nuclear stain with the pSTAT5 signal in CD25+ T lymphocytes (yellow) as compared to all lymphocytes (orange) gated on FoxP3 versus pSTAT5 dot plot are shown on the left. The higher the Similarity score is, the more the translocation is visualized in the example images of cells shown on the right: pSTAT5 (green) is specifically localized to the nucleus stained with 7-AAD (red) in CD25+ T lymphocytes. (b) pSTAT5 response to IL-2 stimulation in whole blood CD4+ Th lymphocytes and their FoxP3+ and FoxP3− subsets from healthy donors was assayed by measuring median fluorescence intensities (MFI) and represented as a ratio of induction to baseline levels (MFI fold change = MFI cytokine-stimulated/MFI unstimulated cells). Box and whisker plots of pSTAT5 response after IL-2 stimulation in FoxP3hi expressing aTregs as compared to FoxP3lo and FoxP3− Th subsets from healthy donors (n = 7). (c) Box and whisker plots of STAT1 protein expression in FoxP3hi expressing aTregs as compared to FoxP3lo expressing Th subsets from healthy donors (n = 6). P values were calculated using the Wilcoxon matched-pairs signed-rank test. ∗ P < 0.05; ∗∗ P < 0.01.