Fig 3.

Analysis of all datasets reveals commonalities in the effects of Tregs and hypoxia on the activated CD8⁺T cell surfaceome.A, left, Venn diagram showing intersection of proteins that were upregulated upon CD8⁺ activation, but downregulated by Treg co-culture or hypoxia. The five proteins at the intersection of all three datasets are indicated. A, right, Venn diagram showing intersection of proteins downregulated upon CD8⁺ activation, but upregulated by Treg co-culture or hypoxia. B, Western blot for SLC5A6, SLC7A1, and SLC16A3. CD8⁺ T cells from three different donors were activated with anti-CD3/anti-CD28 beads in either normoxic (20% O₂, “N”) or hypoxic (1% O₂, “H”) conditions for 3 days before samples were collected for Western blot. A total protein stain is shown as a loading control. C, flow cytometry of cells from N = 7 donors were either left resting or stimulated in normoxic (20% O₂) or hypoxic (1% O₂) conditions for 3 days. Cells were then collected and stained for either IL18R1 or CD70. Mean fluorescence intensity (MFI) values are shown for each donor. Bars represent means and error bars show standard error of the mean. Statistical significance was determined using a one-way ANOVA with a Tukey’s multiple comparisons test (∗ indicates p < 0.05). Treg, regulatory T cell.