Fig. 6
AIL blocked Treg differentiation through targeting c-Jun. A-E Naive CD4 + T cells were isolated from the spleen of C57BL/6 mice, seeded at a density of 1 × 106 cells per well in 24-well plates, and finally cultured with AIL-treated B16-F10 supernatant or AIL-treated YUMM1.7 supernatant under Treg-prone conditions. After 3 days, CD25 + Foxp3 + cells were detected by flow cytometry, and Foxp3 mRNA levels were determined by qRT–PCR. (A) Schematic of the treatment plan. B-C Relative quantification of Tregs differentiated from naive CD4 + T cells. D-E Foxp3 mRNA level. F-I Naive CD4 + T cells were isolated from the spleen of C57BL/6 mice, seeded at a density of 1 × 106 cells per well in 24-well plates, and finally cultured with AIL-treated vector/c-Jun-overexpressing B16-F10 supernatant or AIL-treated vector/c-Jun-overexpressing YUMM1.7 supernatant under Treg-prone conditions. After 3 days, CD25 + Foxp3 + cells were detected by flow cytometry, and Foxp3 mRNA levels were determined by qRT–PCR. F-G Representative flow cytometric plots and relative quantification of Tregs differentiated from naive CD4 + T cells. H-I Foxp3 mRNA level. Data are shown as the mean ± SD, n = 3; *p < 0.05, ** p < 0.01 and *** p < 0.001