Fig. 3. Dapl1 expression marks a distinct subpopulation of naïve CD8 T cells.

(A) Representative UMAP projection of multicolor flow cytometry data from splenic T cells of Dapl1^ZsG/WTFoxp3^RFP/RFP mice. (B) CD122 and CD44 expression on ZsG⁻ or ZsG⁺ splenocytes (left), with summary statistics (right). (C) Surface marker expression on naïve (CD44^lowCD62L^high) T cells, calculated either as the proportion of Ly6C⁺ cells or MFI relative to the ZsG⁻ population (CD5, CD44, CD62L, and CD127). (D) ZsG expression among circulating WT Dapl1^ZsG/WT, Dapl1^ZsG/WTP14⁺Rag2^−/−, or Dapl1^ZsG/WTOT-I⁺Rag2^−/− CD8 T cells. (E) ZsG expression in CD45.2⁺ CD8 T cells 30 days post–adoptive transfer of sorted ZsG⁻ or ZsG⁺ CD45.2⁺ CD8 T cells into CD45.1 hosts. (F) scRNA-seq analysis of naïve CD8 T cells from P14⁺Rag2^−/−, OT-I⁺Rag2^−/−, or WT datasets (GSE131847, GSE199563, GSE213470, GSE221969, GSE181784, and GSE186839), consisting of (top) violin plots representing Dapl1 heterogeneity, (middle) descriptions of the datasets, and (bottom) a heatmap of DEGs consistent among datasets. FC, fold change. (G) Pathway enrichment analysis denoting significantly different pathways between Dapl1⁺ and Dapl1⁻ naïve CD8 T cells. JAK, Janus kinase; STAT, signal transducers and activators of transcription. Statistical analysis for (B) and (C) was calculated via two-tailed paired t tests for difference [(B) and (C)] or two-way ANOVA (E). All data are pooled from three biological replicates from at least two independent experiments [(A) to (C)], or representative of at least three independent experiments [(D) and (G)].