MYC, E2F targets, and cell cycle–related gene upregulation in PBL. (A) Volcano plot showing differentially expressed genes in PBL vs control (log₂ fold change [log₂FC] > |1|; adjusted P [P.adj] < .05), with significantly upregulated genes highlighted in red (1619 genes) and downregulated genes in blue (1337 genes). Log₂FC is depicted in the x-axis and log₁₀(Padj) in the y-axis. (B) GSEA of hallmark data set for upregulated genes in PBL. The figure includes different comparisons between PBL and controls depending on the presence of EBV and MYC-t in tumoral cells. Both, NES and Padj are represented by color and symbol size, respectively. (C) GO analysis for downregulated genes in PBL. The figure shows NES (x-axis) and the gene sets with Padj <.0001 (y-axis). (D) Infiltrating immune cells (%) in PBL according to CIBERSORT immune deconvolution analysis. (E) Heat map displaying the expression of upregulated E2F targets in PBL, involved in the cell cycle. PBL and controls (top) are identified in purple and pink, respectively. The color of the heatmap is represented by the z-score. (F) Correlation between the expression of E2F and their 20 targets associated with G2M checkpoint in PBL patients according to hallmark and GO data sets. R value is represented with color heatmap and P value by circle size. (G) GSEA based on hallmark gene set in EBV-positive vs EBV-negative PBL including all coding genes. P value and gene counts are represented by color and symbol size, respectively. (H) Differential expression of the E2F members (top) and the E2F targets BIRC5 and CSKS2 (bottom) depending on EBV infection. Statistical analysis was performed using Spearman test for correlation analysis in panel F; Dunn multiple comparisons test in panel H; ∗P < .05; ∗∗P < .01; ∗∗∗P < .001. DC, dendritic cell; GO, gene ontology; GSEA, gene set enrichment analysis; NES, normalized enrichment score; NK, natural killer; ns, not significant; PC, plasma cell; Th, T-helper; TNFA, tumor necrosis factor alpha; Treg, regulatory T cell.