Transcriptomic analysis of LPS-stimulated M-CSF–derived human macrophages after the addition of exogenous Ado and PGE₂reveals a modest but similar change in gene expression. Human monocytes were cultured in M-CSF for 7 days and then left unstimulated, or they were stimulated with LPS, LPS + Ado, or LPS + PGE₂ for 4 hours before total mRNA was isolated and sequenced on an Illumina platform. (A) The top 10 most highly upregulated and bottom 10 most highly downregulated genes by LPS + PGE₂ (light blue bars) and LPS + Ado (light green bars) are expressed as log₂ fold change (log₂FC) relative to LPS stimulation alone. Differentially expressed genes (DEGs) in common between LPS + Ado and LPS + PGE₂ are marked with an asterisk (FC >2; P < .05). Note: CCL3L3 and CCL3L1 share the same ENSEMBL gene ID and were therefore represented together as 1 bar. (B) Venn diagrams indicate the number of unique and shared upregulated (top) and downregulated (bottom) DEGs (FC >2; P < .05) of macrophages stimulated by LPS + Ado and LPS + PGE₂ relative to macrophages stimulated by LPS alone. (C) The log₂FC of genes expressed by LPS + Ado vs LPS + PGE₂ relative to LPS alone are visualized in a scatter plot. The R value calculated by Pearson’s correlation analysis is indicated in the plot area (P < 2.2e-16). (D) Single-cell RNA-seq of unstimulated macrophages or macrophages stimulated by LPS, LPS + Ado, or LPS + PGE₂ at 4 hours is visualized by using uniform manifold approximation and projection (UMAP). (E) The top 20 and bottom 20 DEGs (FC >2; P < .05) that are shared between macrophages stimulated by LPS + Ado (light green bars) and LPS + PGE₂ (light blue bars) are expressed as the FC relative to macrophages stimulated by LPS alone. Genes with published roles in growth promotion and angiogenesis are highlighted in lavender; genes with published roles in inflammation are highlighted in red. (F) The top 5 enriched molecular function Gene Ontology (GO) terms for the DEGs shared between LPS + Ado and LPS + PGE₂ (FC >2; P < .05) are indicated. Point size corresponds to the number of DEGs in each GO category. The color of the point indicates the P value of enrichment for each GO category. The rich factor represents the ratio of the number of DEGs to the number of genes in the GO term category.