Fig. 2.

TRE serum induces widespread transcriptional reprogramming in endothelial cells. A Volcano plot showing differentially expressed genes (DEGs) in hCMEC/D3 endothelial cells following treatment with TRE serum compared to aged control serum. Genes are plotted by log₂ fold change and –log₁₀ adjusted p-value. Significantly upregulated genes are shown in red and downregulated genes in blue (adjusted p-value  < 0.05, |log₂FC|> 1). The stress-responsive gene GDF15 is highlighted as a representative strongly induced transcript. B Summary of the number of significantly differentially expressed genes, demonstrating a predominance of upregulated transcripts in response to TRE serum. C Heatmap of the top differentially expressed genes with hierarchical clustering. Samples cluster according to treatment condition (control vs. TRE), with distinct transcriptional profiles characterized by coordinated upregulation (red) and downregulation (blue) of gene sets, indicating a robust and consistent treatment effect. D Gene Ontology (GO) biological process enrichment analysis of upregulated genes. The most significantly enriched pathways include cell communication, signaling, response to stimulus, angiogenesis-related processes, and vascular development, suggesting activation of coordinated stress-responsive and adaptive signaling programs in endothelial cells exposed to TRE serum