Fig. 4

Cytokines secretion profile evaluation of T_conv cells induced by trophoblast cells-derived IL-35 and the related transcription factors which mediate the conversion. a T_conv cells were cultured in medium (control) alone, or with human r-sc-IL-35 (IL-35) (50 ng ml⁻¹), or with trophoblast cells supernatant (with or without neutralizing anti-IL-35 mAb) for 5 days. Then the supernatants of activated T_conv cells were collected for cytokines secretion profile evaluation using Bio-Plex Protein Array system. In T_conv cells co-cultured with supernatant from PT or HTR8 cells, final results of cytokines concentration were normalized to the initial supernatant from PT or HTR8 cells, respectively, which were used as inner control (n = 2). b T_conv cells were cultured in medium (control), or with human r-sc-IL-35 (IL-35) (50 ng ml⁻¹ or 100 ng ml⁻¹), or with trophoblast cells (PT and HTR8) supernatant for 1 h. Then, T_conv cells were collected and the protein levels of phosphorylated (p) and total STAT1, STAT3, and STAT4 were determined by western blot (n = 3). Data were represented as mean ± SD. Significance was determined by Student’s t-test. *P < 0.05 and **P < 0.01