Fig. 4. Compound 182 represses MC38 and AT3 tumor growth.

a–c MC38 colon tumor cells were xenografted into the flanks of 8-week-old C57BL/6 male mice. Mice were treated with Compound 182 (CMP-182; 10 mg/kg i.v.; n = 5) or saline (n = 7) on days (d) 7, 9, 11, 13, 15, 17, and 19 after tumor cell implantation. a Tumor growth was monitored and tumor weights were measured. b, c Tumor-infiltrating lymphocytes (TILs) including CD44^hiCD62L^lo CD8⁺ and CD4⁺ effector/memory (EM) T cells, CD19⁺ B cells, NK1.1⁺TCRβ⁻ (NK) cells, CD4⁺CD25⁺FoxP3⁺ regulatory T cells (T_regs) and granulocytic and monocytic CD11b⁺F4/80^hi/loLy6C⁺Ly6G^+/− myeloid-derived suppressor cells (MDSCs) were analyzed by flow cytometry. In (b) intracellular granzyme B (GRZMB) and cell surface PD-1 and TIM-3 were detected in unstimulated tumor-infiltrating CD8⁺ T cells. d–f AT3 mammary tumor cells were injected into the fourth inguinal mammary fat pads of 8-week-old C57BL/6 female mice. Mice were treated with CMP-182 (10 mg/kg i.v.; n = 6) or saline (n = 8) on days (d) 10, 12, 14, 16, 18, 20, 22, 24, and 26 after tumor cell implantation. d Tumor growth was monitored and tumor weights measured. e, f TILs (CMP-182: n = 5; Saline: n = 7) including CD4⁺ EM T cells, CD19⁺ B cells, NK cells, T_regs and MDSCs were analyzed by flow cytometry. In (e) intracellular GRZMB and cell surface PD-1 and TIM-3 were detected in unstimulated tumor-infiltrating CD8⁺ T cells. In (a–f) representative results (means ± SEM) from at least two independent experiments are shown. Significance for tumor sizes in (a, d) was determined using a 2-way ANOVA Test and for tumor weights in (a, d) using a 2-tailed Mann–Whitney U Test. In (b, c, e, f) significances were determined using a 2-tailed Mann–Whitney U Test.