FIGURE 4.

Effect of the conditioned media from enEVTs on the differentiation of Tregs. (A‐E) The results of flow cytometry showing the influence of the conditioned media from enEVTs (enEVT‐CM) on the differentiation of human Tregs. Human peripheral naïve CD4⁺ T cells are isolated and cultured in cell‐free RPMI‐1640 medium (CTRL; A), 50% RPMI‐1640 medium + 50% normal enEVT‐CM (B), 50% complete RPMI‐1640 medium + 50% normal enEVT‐CM + 20 μg/mL normal mouse IgG (C), 50% complete RPMI‐1640 medium + 50% normal enEVT‐CM + 20 μg/mL blocking antibody against TGF‐β1 (D), or 50% RPMI‐1640 medium + 50% RSA enEVT‐CM (E). The proportions of CD4⁺ CD25⁺ FOXP3⁺ Tregs were analysed after culture. F, Statistical analysis of flow cytometry showing the proportions of human CD4⁺ CD25⁺ FOXP3⁺ Tregs upon various treatments. The statistical analysis was performed based on the results from three independently repeated experiments using different batches of enEVT and naïve CD4⁺ T cells. Data are presented as mean ± SD, and the comparisons between groups were finally accomplished with Student's t test. *P < .05