Potent inflammatory state induced in the tumor microenvironment. Mice treated with 11 mg/kg 1V270-micelles and evaluated after 24 h by Nanostring in A and after 6 and 24 h by ELISA in B and by flow cytometry in C and E–J. (A) Gene expression in CT26 tumors. Shown are genes in the cytokine activity gene-ontology annotation (GO: 0005125) with a Benjamin-Yekutieli adjusted p value between untreated and 1V270-micelle treated below 0.05 (n = 5). Blue are >2-fold upregulated, red are >2-fold downregulated genes. (B) Cytokine levels in CT26 tumors and spleens (n = 4–5). (C) Erythrocytes in CT26 tumors (n = 4–6). (D) Representative macroscopic pictures of CT26 tumors. (E) Viability in spleens, liver and CT26 tumors (n = 4–6). (F) t-SNE visualization of viable cells in CT26 tumors with populations colored. Mean cellular density of each group is overlaid in the t-SNE visualizations. (G) Cancer cells in CT26 tumors. (H) Immune cells in CT26 tumors. (I) Neutrophils in CT26 tumors. (J) CD69 expression on effector cells in CT26 tumors. Gating for tumor data in C, E, and G is shown in Fig. S8 and for liver and spleen data in E in Fig. S9. Gating for F and H–J is shown in Fig. S10. Statistical analyses were Kruskal-Wallis with Dunn’s correction (B, C and H, I) and 2-way ANOVA with Dunett’s multiple comparison tests (E and J). MFI, median fluorescent intensity.