Figure S3 from Protumor Macrophage Polarization Mediated by BMP2/4–ACVR1 Signaling Orchestrates an Immunosuppressive Microenvironment during Tumor Initiation

Extended Data Fig.3. Macrophages coordinate tumor microenvironment and sharply change during tumor formation, related to Figure 2 (A) Network depicting the interactions occurring among various cell types at each lung cancer stage. Each node is a cell type. Each color indicates each cell type. The arrow width is the sum of interaction values between two clusters. The results were both from dsCellnet(up) and cellchat(bottom). (B) The number of ligands (left) and receptors (right) number in each cell type at each lung cancer stage, compared to control lungs. Each color represents a cell type. (C) “Betweenness scores” of each cell type in the cell–cell communication network in Figure 3A, indicating that macrophages apparently function as central mediators during tumorigenesis. (Betweenness score measures the number of times a node lies on the shortest path between other nodes.) (D) H&E staining images of lung sections from mice fed without doxycycline(left), with doxycycline at 2 week(middle) and with doxycycline and lipo(right). scale bar :50um. (E) MRI imaging of mouse lungs at 4 weeks after tumor induction, following intranasal administration of clodronate liposomes at week 2 to deplete macrophages. The treated mice exhibited significant tumor suppression as visualized by MRI. (F) Expression levels of selected marker genes in the different subtypes of monocytes and macrophages in t-SNE projection. Each color indicates one cell subtype. (G) Heatmap showing the expression patterns of the top 10 ranked marker genes for each monocyte and macrophage sub cluster. (H) tSNE projection of single cells at each time point from the two biological replicates, highlighting the high repeatability of each macrophage compositions in the two experiments. (I) Bar plot quantifying the proportions of various macrophages at each time point from the two biological replicates, highlighting the high repeatability of macrophage compositions in the two experiments. (J) The G2M score of Mac subclusters. Each color represents a cell type. (K)Enrichment analysis of up-regulated genes from Mac_a1_vs_Mac_b and Mac_a2_vs_Mac_c.