Fig 1: Th17 cultures containing IL‐21+ TGF‐β1 (Th17.d) develop independently of T cell–intrinsic IL‐6. WT or Il6 KO CD4+ T cells were differentiated under Th17.d conditions for 7 days, and Il17A expression was assessed by ELISA (A) and qPCR (B). Responsiveness to CD8+ T‐cell‐mediated suppression was evaluated in coculture assays (C). In parallel, WT cells were cultured with or without anti–IL6R antibody and subjected to the same functional readouts (D–F). Data are from three independent experiments (n = 6 per group); each point represents an individual donor. Error bars indicate SEM. Statistical analysis was performed using paired or unpaired Student's t‐test (*p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001; **p ≤ 0.0001).
Fig 2: Schematic diagram outlining the overall experimental approach. (A) PBMCs were isolated from healthy donor leukoreduction cones, and naïve CD4+ T cells were purified by magnetic separation. (B) Naïve CD4+ T cells were used for CRISPR‐Cas9 knockout experiments with gene‐specific gRNAs, and KO efficiency was assessed after nucleofection. (C) Cells were then subjected to Th17 differentiation under four cytokine conditions (Th17.a, Th17.b, Th17.c, and Th17.d) in different combinations of TGF‐β1, IL‐6, IL‐1β, or IL‐21, with anti‐CD3/CD28 activation for 7 days.
Supplier Page from Cell Sciences for Recombinant Human IL-21
Protein/Peptide:IL-21
Protein/Peptide Species:Human