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. 2019 Jul 12;10:1619. doi: 10.3389/fimmu.2019.01619

Figure 5.

Figure 5

Cytokine and mRNA expression analysis. Naïve CD4+ T cells (Naïve), CD25−LAG3+ T cells (CD25−LAG3+), CD25+LAG3+ T cells (CD25+LAG3+), activated CD25+ Tregs and CD4+CD25−LAG3−CD45RA− T cells (Memory) donated by HC were analyzed. (A) Representative findings of intracellular staining. CD4+ T cell subsets were separated by sorting and stimulated for 72 h with anti-CD3ε mAb (10 μg/mL) and anti-human CD28 mAb (5 μg/mL) in the presence of recombinant human IL-2 (100 IU/mL). (B) Summary of intracellular staining of CD4+ T cell subsets taken from three HC individuals. *P < 0.05; **P < 0.01; ***P < 0.001. One-way ANOVA followed by Tukey's multiple comparisons test. (C) Relative mRNA expression of FOXP3 compared to GAPDH in sorted CD4+ T cell subsets (n = 3) without stimulation. Relative mRNA expression of RORC compared to GAPDH in sorted CD4+ T cell subsets (n = 3) stimulated for 72 h with anti-CD3ε mAb (10 μg/mL) and anti-human CD28 mAb (5 μg/mL) in the presence of recombinant human IL-2 (100 IU/mL). Cell subsets were **P < 0.01; ***P < 0.001; ****P < 0.0001. One-way ANOVA followed by Tukey's multiple comparisons test.