Fig. 1

Identifying Tregs and Tresps and analysis of FOXP3 expression in isolated CD4⁺CD25^highCD127^low/− T cells. We immunolabeled PBMCs with PerCP-Cy5.5-labeled anti-CD4, phycoerythrin-labeled anti-CD25, and Alexa Fluor® 647-labeled anti-CD127. a CD4⁺ T cells were gated as CD25^high, CD25^low, and CD25⁻ based on the level of CD25 expression in CD4⁻ T cells. CD4⁺CD25⁻ cells were isolated as Tresps. b We identified Tregs as CD25^highCD127^low/− cells among the CD4⁺ T cell population. c To verify the purity of the sorted Tregs, we stained the cells with Alexa Fluor® 488-labeled anti-FOXP3. The percentage of FOXP3 expression in freshly isolated CD4⁺CD25^highCD127^low/− T cells is shown. The data revealed that sorted Tregs were > 95% positive for FOXP3 in URSA patients (n = 12) and normal controls (n = 12). Representative data of a patient with URSA (left panel) and a normal control subject right panel) are shown