Figure S3.

Effects of coculture with WT or CD25-deficient TRCs on survival, proliferation, and activation of CD4⁺ T cells. Purified, CFSE-labeled naive CD4⁺ T cells (2 × 10⁵) were cultured without or with either WT or CD25-deficient TRCs (2 × 10⁴) in the presence or absence of soluble anti-CD3 and CD28 antibodies (3 and 1 μg/ml) with irradiated APCs (2 × 10⁵) for 72 h. (A) Cell viability was assessed by FACS analysis with viability dye staining. ***, P < 0.001 compared with naive CD4⁺ T cells alone. Error bars indicate the mean ± SEM (n ≥ 5 per group). (B) The proliferation of CFSE-labeled CD4⁺ T cells was measured by FACS analysis. (C) Representative flow cytometric analysis of FSC/SSC and CD4⁺/CD69⁺ cell profiles. Data are representative of three independent experiments.