Purification control of CD4+ Th cells and CD4+CD25+ nTreg cells by FACS analysis. CD4+ Th cells and nTregs were isolated from human leukapheresis (up to 1.5×1010 whole PBMC cells) of healthy volunteers. Populations were stained with the indicated markers using fluorescence-labeled mAbs and analyzed by flow cytometry. The figure shows the dot blot analysis of ungated cell measurement. The cells were stained with CD4 APC (BD Biosciences), CD25 PE-Cy5 (BD Bioscience), CD25 PE-Cy7 (BD Biosciences), CD8 PerCp-Cy5.5 (BD Biosciences) and CD19 APC-Cy7 (BD Biosciences), CD3 FITC/CD56/CD16 PE (BD Bioscience) or CD14 FITC (BD Biosciences) and analysed by FACS CantoII (BD Bioscience). The purity of the resulted nTregs was typically >94% and for CD4+ Th cells >98%. (0.89 MB TIF)