Figure 8. HIF stabilization partially reproduces hypoxia-induced T_RM phenotype.

Naive CD8⁺ T cells were activated in 20% O₂ (AtmosO₂) in the presence of FG-4592 for 4 days and then for an additional 2 days with TGF-β1. Cells activated in 2% O₂ with addition of TGF-β1 on day 4 are shown in red for comparison. (A) Frequency of the CD69⁺CD103⁺ population and (B) expression of T_RM-associated markers by the CD69⁺CD103⁺ population were assessed by flow cytometry, with representative results shown in A for 1 donor, n = 3. Repeated measures 1-way ANOVA (A and B) followed by test for trend (A) or Dunnett’s multiple comparisons test (B), *P adj < 0.05, ***P adj < 0.001; data are mean ± SEM. CD69⁺CD103⁺ cells generated in FG-4592 or 2% O₂, with addition of TGF-β1 were sorted before RNA isolation and analysis via mRNA sequencing (n = 2–3), with differential expression determined by |log₂FC| ≥ 1 and FDR < 0.05. (C) Venn diagram showing the overlap of DEGs between CD69⁺CD103⁺ cells generated via FG-4592 or 2% O₂ relative to 20% O₂ (all with addition of TGF-β1). (D) Normalized expression levels of selected DEGs between CD69⁺CD103⁺ cells generated in FG-4592 or 2% O₂, showing min, median, and max; ****P adj < 0.0001. CD69⁺CD103⁺ cells generated via TGF-β1 alone (20% O₂) are shown for reference.