Fig. 8

RNA-seq assay to investigate the immune profile of the B16 tumors. a The heatmap presenting the relative expression levels of differentially expressed genes (n = 4 biological replicates). b–e Gene set enrichment analysis (GSEA) showing the significantly enriched biological processes in Mn²⁺ plus YM101-treated tumors, relative to YM101-treated tumors. The biological processes including activation of immune response, activation of innate immune response, positive regulation of adaptive immune response, IFN-I production, IFN-I signaling pathway, response to IFN-I, T cell activation, NK activation, B cell activation, macrophage activation, myeloid dendritic cell activation, cell killing, T cell mediated cytotoxicity, NK mediated cytotoxicity, and antigen processing and presentation were significantly enriched in the combination group. f–k The scores of signatures of T cell, NK, dendritic cell (DC), macrophage, IFN-α response, and IFN-γ response. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 denote the significant difference relative to combination treatment