Figure 2.

Gene expression profile shows reduced exhaustion and increased memory genes in CD8 T cells treated with MDM2 inhibitor. (A) Principal component analysis (PCA) of bulk RNA-seq between untreated (purple) and NAVtreated (orange) CD8+ T-cell samples when grouped based on overall transcriptomic similarity (n = 3 biological replicates). Hierarchical clustering heatmap of 6,313 differentially expressed genes (|log₂FoldChange| > 1, padj < 0.01). Rows represent genes and columns represent biological replicates. Red and blue indicate relative upregulation and downregulation (z-score scaled expression), respectively. (B) Volcano plot shows differential gene expression (DGE) between untreated and NAV-treated groups. Each dot represents a significantly upregulated gene in the group as shown (|log₂FoldChange| > 1, padj < 0.01). Key memoryassociated and effector-associated genes (purple) are labeled. Bar plot of log₂ fold change shows top upregulated genes within each group. (C) GSEA shows NAV-treated cells have significant positive enrichment of genes upregulated in memory and significant negative enrichment of genes upregulated in effector. Normalized enrichment score (NES), running enrichment score (green line), and ranked genes (barcode) are shown. Genes ranked closest to the left (rank = 1) are most upregulated in NAV-treated cells, while those furthest to the right are most upregulated in untreated cells. Adjusted p value of less than 0.05 was deemed significant. (D) CirGO visualization of enriched Gene Ontology (GO) biological process terms for differentially expressed genes. Inner rings display higher order GO categories, and outer rings show individual terms.