Fig. 5

Macrophage ACT activated inflammatory immune responses in the TME. (A) UMAP analysis of Cd45 + sorted tumor infiltrated immune cells from Control and M1 + NextA macrophage treated tumors analyzed with 10xGenomics workflow. UMAP cluster analysis of aggregated immune cells by SingleR cell annotation revealed major immune cell populations. (B) Tumor macrophage subclusters Mac1, Mac2, Mac3, and Mac4 show differential presence represented as a percentage of total tumor macrophages (insert with stacked bar plot). (C) Feature plots representing the expression of inflammatory genes Cxcl9 and Cd72 associated with M1 phenotype in Mac1 macrophage subcluster in Control and M1 + NextA tumors. (D) Feature plots representing expression of tumor promoting genes Arg1 and Mrc1 associated with M2 phenotype in Mac2 macrophage subcluster in Control and M1 + NextA tumors. (E) Bubble plot showing differentially expressed top significant gene between macrophage subcluster. Mac1 associated genes are shown in orange, and Mac2 associated genes are shown in green color. (F) Heatmap representing the expression of Mac1 and Mac2 gene signatures in single cells across 4 macrophage subclusters. (G) Circos plot showing potential interactions between ligands expressed by macrophages in Mac1 and Mac2 subclusters and receptors expressed on the receiver T-cells. Heatmap representing the pathway analysis of activated target genes in receiver monocytes. MSigDB Hallmark 2020 module in Enrichr was used to generate the heatmap. (H) Circos plot showing potential interactions between ligands expressed by macrophages in Mac1 and Mac2 subclusters and receptors expressed on the receiver monocytes and corresponding heatmap of pathway analysis