Figure 3. TSLP modulates gene expression on naïve and memory CD8⁺ T cells in vitro.

RNA-Seq performed on sorted naïve and memory CD8⁺ T cells after 4 or 24 hr incubation with medium or TSLP with or without TCR stimulation. Shown are the number of differentially expressed genes (FC >1.5, FDR < 0.05). (A and B) Number of genes affected by TSLP in CD8⁺ T cells not stimulated (A) or stimulated (B) with anti-CD3 + anti-CD28. (C) Venn diagram showing the number of genes whose expression was upregulated or downregulated by TSLP without TCR stimulation at 24 hr of incubation in naïve and memory CD8⁺ T cells. (D) Genes regulated by TSLP both in naïve and memory CD8⁺ T cells, color-coded as indicated in the text. (E) Genes differentially regulated by TSLP in naïve vs. memory cells. Highlighted are those that were selectively induced or repressed after stimulation with TSLP for 24 hr.

Figure 3—figure supplement 1. Gating strategy for sorting memory CD8⁺ T cells for in vitro stimulation.

Shown is forward versus side scatter (FSC versus SSC) to exclude debris (first plot), then FSC-A and FSC-H analysis as well as SSC-H and SCC-W were performed to exclude doublets (second and third plots). Then dead cells were eliminated (fourth plot). Transferred P14 cells were identified as CD8⁺ Thy1.1⁺ cells (fifth plot).