Extended Data Fig. 2. Sucralose limits T cell proliferation without affecting viability.

a) CD4⁺ T cell proliferation assay performed with αCD3 (5 μg ml⁻¹) and αCD28 (1 μg ml⁻¹) for 3 days in the presence of serial dilutions of the indicated sweeteners. b) Proliferation of CFSE stained CD8⁺ T cells exposed to 0.5 mM of the indicated sweeteners and activated with a high dose of αCD3 (5 μg ml⁻¹) or low dose of αCD3 (1 μg ml⁻¹) in the presence or absence of αCD28 (1 μg ml⁻¹). c) Jurkat T cell proliferation in control media or media supplemented with the indicated sweeteners. N = 3. d) Percentage of viable CD8⁺ (left) and CD4⁺ (right) T cells 24 h post activation with αCD3 (2 μg ml⁻¹) and αCD28 (1 μg ml⁻¹). Cell viability was determined by Fixable Viability Dye eFluor780® exclusion. Means represent ± s.e.m. for biological (d) or technical (c) replicates. Statistical significance was determined using one-way ANOVA with Tukey’s (c) or Dunnet’s (d) multiple comparison test. Data are representative of 3 independent experiments (c).

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