Fig. 6: Tregs promote Th17 cell differentiation by elevating the p-STAT3/p-STAT5 ratio in Teffs.

A–D Naïve CD4⁺ T cells isolated from C57BL/6 mice were stimulated with anti-CD3 and APCs in the presence or absence of Tregs (1:1 ratio), and cultured in the indicated polarization conditions. Representative flow cytometry plots and bar graphs showing frequencies of p-STAT5⁺CD4⁺ T cells (A, B) and p-STAT3⁺CD4⁺ T cells (C) in T cells cultured for 24 hours. D Bar graph showing the ratio of p-STAT3⁺CD4⁺cells to p-STAT5⁺CD4⁺cells. E–H Naïve CD4⁺ T cells isolated from C57BL/6 mice were cultured with anti-CD3 and anti-CD28 in the indicated polarization conditions. E Immunoblotting of STAT5, p-STAT5, STAT3, and p-STAT3 in T cells cultured for 24 h. F Density values of the Immunoblot were measured using Image J, and the ratio of p-STAT3 to p-STAT5 was calculated. The samples derive from the same experiment, and the blots were processed in parallel. G Representative flow cytometry plots showing frequencies of p-STAT5⁺CD4⁺ T cells after 24 h culture. H Bar graph showing the ratio of p-STAT3⁺CD4⁺ T cells to p-STAT5⁺CD4⁺ T cells after 24 h culture. Data are pooled from three biological replicates. Data were analyzed by unpaired two-tailed Student’s t tests (B, C and D) or one-way ANOVA with Tukey’s post hoc test (F and H). Summary data are presented as mean ± SEM, ns, not significant. Source data are provided as a Source Data file.