Figure 4

Foxp3 expression in T cells after co-culture was detected by RT-PCR, Real-time PCR or Western blot. (A) Analysis of RT-PCR products of Foxp3 and comparison of the gray scale value between Foxp3 and β-actin by agarose gel electrophoresis. Three separate experiments were carried out. RT-PCR product of β-actin and Foxp3 from the total mRNA isolated from CD3⁺T cells cultured with growth medium, or from the T cells co-cultured with IDO gene-transfected CHO cells, or from the T cells co-cultured with CHO/EGFP cells. The value of the Foxp3 and β-actin gray scale ratio in T cells after 7 days of co-culture with IDO gene-transfected CHO cells was higher than in the control groups (P < 0.05). (B) Expression of Foxp3 gene analyzed by real-time RT-PCR. Three separate experiments were carried out. Amplification curve of Foxp3 in the IDO gene-transfected group and the control groups. Expression of Foxp3 in T cells after 7 days of co-culture with IDO gene-transfected CHO cells was higher than that in the control groups (P < 0.05). (C) Expression of Foxp3 analyzed by Western blot analysis. Three separate experiments were carried out. Expression of Foxp3 protein in the CD3⁺T cells cultured with growth medium for 7 days; or 7 days after co-culture with CHO/EGFP cells; or 7 days after co-culture with IDO⁺ CHO cells. No Foxp3 protein was detected in the control groups.