Fig. 2

Detection of cell surface molecules and signaling pathway molecules after knockdown and over-expression of Foxo1 in Treg cells. a, Representative expression of the Foxo1, CD127, CD103, ICOS, Foxp3 or CD25 in Treg cells 48 h after transfection with Foxo1 siRNA by flow cytometry (broken black line: isotype, green line: control, red line: Foxo1 siRNA). b Representative expression of the Foxo1, CD127, CD103, ICOS, Foxp3 or CD25 in Treg cells 48 h after transfection with over-expression plasmid of Foxo1 by flow cytometry (broken black line: isotype, green line: control, red line: Foxo1 over-expression). c Detected of Median Fluorescence Intensity (MFI) for CD127 in Treg cells 48 h after transfection with Foxo1 siRNA and Foxo1 over-expression plasmid by flow cytometry. d Representative western blot of p-Erk1/2, total Erk1/2, p-Akt, total Akt, p-Stat5, total Stat5, p-Foxo1 and total Foxo1 in Treg cells 48 h after transfection with Foxo1 siRNA and over-expression plasmid of Foxo1, GADPH was used as a control. Treg cells stimulated with anti-CD3 (0.01 μg/ml) and anti-CD28 (1.0 μg/ml) in medium during culture. e Expression of mRNA for IL-2, IL-4, IL-7 and IL-15 in Treg cells, 48 h after transfection with Foxo1 siRNA and Foxo1 over-expression plasmid. All experiments were repeated at least three times. **P < 0.005, n.s: no significance