Analysis of Mif expression by scRNA-seq. (A) UMAP of human neuroblastoma cells, clustered and colored by cell type. (B) Violin plot presenting MIF expression in the different clusters of A. (C) UMAP of TH-MYCN mouse NB cells, clustered and colored by cell type. (D) Violin plot presenting Mif expression in the different clusters of C. (E) UMAP of selected macrophage cells reclustered from C. (F) Heatmap of correlation scores between TH-MYCN TAMs and Mycn-nGEMM TAMs subtypes. Columns represent Mycn-nGEMM TAMs and rows represent TH-MYCN TAMs subtypes. The colored bar at the top represents the Mycn-nGEMM macrophage clusters (subtypes). (G) Levels of N-Myc protein in Mycn-nGEMM-high, Mycn-nGEMM-low, and TH-MYCN^+/− tumors, assessed by immunostaining with Cell Signaling Technologies antibody against N-Myc. Scale bar = 50 µm. (H) Representative BLI images from experiment described in Fig. 5 A. (I) Growth curves of Mycn-nGEMM-high tumors treated with IgG, anti–PD-L1, and CD40 agonist of the combination, as assessed by BLI. Grey area indicates treatment window. Data represent mean values ± SEM (n = 5 mice per group). (J) Kaplan–Meier curves for C57BL/6J mice injected with Mycn-nGEMM-high cell line in the renal capsule and treated with IgG, anti-PD-1, and CD40 agonist of the combination (n = 5 mice per group). (K) Gating strategy for data represented in Fig. 6, E–K. Kaplan–Meier survival analyses were performed by log-rank test (I). ns = no significance.