Figure S2 from Protumor Macrophage Polarization Mediated by BMP2/4–ACVR1 Signaling Orchestrates an Immunosuppressive Microenvironment during Tumor Initiation

Extended Data Fig.2. EGFR driven lung adenocarcinoma resembles a “cold” TME, Related to Figure 1 (A) Gating strategy for living cells in a single cell suspension prepared from a lung tumor, after excluding erythrocytes. (B) Heatmap showing the expression patterns of the top 10 ranked marker genes for each cell type. (C) Bar plot quantifying the proportions of various types of non-immune or immune cells at each time point from the two biological replicates, highlighting the high repeatability of each cell type compositions in the two experiments. (D) The ratio of fibroblasts, lymphatic endothelial cells, endothelial cells, epithelial cells in each induced time point from the two biological replicates, highlighting the high repeatability of each cell type compositions in the two experiments. The dashed line indicates replicate1, Solid line indicates replicate 2. (E-F) Flow cytometry analysis of lung cell suspensions derived from doxycycline-induced mice (2mg/ml) for 0, 1, 2, and 4 weeks. (E) Flow cytometry plots of epithelial cells (CD45-CD326+) (top) and macrophages (CD45+Ly6G-CD64+) and neutrophils (CD45+Ly6G+) (bottom). (F) Flow cytometry frequencies of neutrophils (CD45+Ly6G+) (top left), monocytes (CD45+Ly6G-MHCII-CD64-CD11C-Siglecf-CD11b+Ly6C+) (top right), B cells (CD45+CD19+) (bottom left), and T cells (CD45+CD3+) (bottom right) at the indicated time points. Each dot represents an individual mouse. (G) Flow cytometry analysis of the absolute number of B cells (CD45⁺CD19⁺, top) and T cells (CD45⁺CD3⁺, bottom) in lung tissue from doxycycline-induced mice (2 mg/mL) at 0, 4, and 8 weeks. Cell counts are shown per milligram of lung tissue, representing cell density-normalized absolute numbers.