Comparative Study of Regulatory T Cell Function of Human CD25⁺CD4⁺ T Cells from Thymocytes, Cord Blood, and Adult Peripheral Blood

Table 1

Combinations of antibodies used in FACS analysis.

	Fluorophore	FITC	PE	PE-TR	PerCP-Cy5.5	PE-Cy-7	APC	APC-Cy7	Pacific Blue
Surface staining^(a)	Ag	CD45RO	CD25	PI	CD8	CD3	CD62L	CD4	CD45RA
	Clone	UCHL1	4E3		SK1	SK7	Dreg56	RPA-T4	MEM-56
Intracellular staining^(b)	Ag	CD62L	CD152	EMA	CD8	CD4	CD25	-	CD45RA
	Clone	Dreg56	BNI3		SK1	SK3	M-A251		MEM-56
	Ag	CD45RA	FOXP3	-	-	CD152	CD25	CD4	CD8
	Clone	HI100	PCH101			BNI3.1	M-A251	RPA-T4	RPA-T8

^(a)All Abs were added to cells except PI, and cells were stained for 20 minutes. PI were added just before FACS analysis.
^(b)All Abs were added to cells except anti-FOXP3-Ab and/or anti-CD152-Ab, and cells were stained for 20 minutes. After cell fixation and permeabilization, anti-FOXP3-Ab and anti-CD152-Ab were added for intracellular staining.