Fig. 4

The JAK–STAT pathway plays a key role in the regulation of Treg/Th17 balance. The Lewis cells were cocultured with SPC and treated with 60 μM CK, 10 nM PTX, or both for 48 h. All experiments were carried out in the co-culture system, including Lewis cells, SPC or the whole system. A, B In addition to CK or PTX, the co-cultured system was cotreated with 10 μM SH-4-54 for 48 h. CD4⁺CD25⁺ cells and CD4⁺CD196⁺ cells in SPC were determined by Flow Cytometry, and the ratio of CD4⁺CD25⁺ cells to CD4⁺CD25⁺ cells was calculated. C In addition to CK or PTX, the co-culture system was cotreated with 10 μM SH-4-54 for 48 h. The concentration of IL-10 and IL-17A in co-culture system was determined by ELISA. D In addition to CK or PTX, the co-culture system was cotreated with or without 10 μM SH-4-54 for 48 h. The expression of RORγ, p-STAT3 and p-STAT5 was detected by Western blot. E, F In addition to CK or PTX, the co-culture system was cotreated with 10 μM SH-4-54 for 48 h. CD3⁺CD4⁺ cells and CD3⁺CD8⁺ cells in SPC were determined by Flow Cytometry, and the ratio of CD3⁺CD4⁺ cells to CD3⁺CD8⁺ cells was calculated. G In addition to CK or PTX, the co-culture system was cotreated with 10 μM SH-4-54 for 48 h. The concentration of IFN-γ in co-culture system was determined by ELISA. Data are mean ± SD for ≥ 3 independent experiments; P values have no significant difference, vs Ctrl