Immune profiling of Mycn-nGEMM tumors reveals a highly immunosuppressive TME dominated by TAMs. (A) t-SNE representation of GFP and CD45 signals in Mycn-high and Mycn-low nGEMM tumors (n = 3 tumors per cell line). (B) Average proportion of tumor cells (GFP⁺), immune cells (CD45⁺), and other cells in Mycn-nGEMM tumors (n = 3 tumors per cell line). (C) Heatmap indicating levels of expression for each marker for all CD45⁺ meta clusters identified with the mouse mass cytometry panel. (D) t-SNE representation of clustering for CD45⁺ cells in Mycn-nGEMM tumors (n = 3 tumors per cell line). (E) Percentage of the different immune subsets in Mycn-nGEMM tumors (n = 3 tumors per cell line) (F) Expression of cytokines assessed by RNA-seq analysis on Mycn-nGEMM-high and Mycn-nGEMM-low cells, represented as normalized read counts. (G) Secreted MIF expression from conditioned cell media of Mycn-nGEMM, MC38 murine colon adenocarcinoma and TCMK1, mouse kidney epithelial cells determined by ELISA assay. Data represent mean ± SD from three independent experiments. Statistical significance was determined by one-way ANOVA multiple comparisons test (G). ***P < 0.001, ns = no significance.