Fig. 2. The immune cell profiles in subcutaneous and liver tumors.

A). The percentages (%) of CD3⁺ cells (CD45⁺ L/D⁻ CD3⁺ NK1.1⁻) in CD45⁺ cells, in subcutaneous tumors (Sub T, left to dotted line) or metastasized liver tumors (Liv T, right to dotted line). The numbers of total CD45⁺ cells in each gram of tumor tissues were first determined and the relative percentages or ratios of various cell types were calculated.

B). The CD8 versus CD4 T cell ratios.

C). The percentages of Treg cells (CD3⁺ NK1.1⁺ CD4⁺ Foxp3⁺) in CD45⁺ cells.

D). The CD8 to Treg cell ratios.

E). The percentages of CD8⁺ GzmB⁺ T cells (CD3⁺ NK1.1⁺ CD8⁺ Granzyme B⁺) in CD45 cells.

F). The percentages of indicated cell subsets in CD8 T cells. Green bar: effector memory T cell (T_em, CD62L⁻CD44⁺); blue bar: naïve T cells (CD62L⁺CD44⁻); red bar, central memory T cells (T_cm, CD62L⁺CD44⁺). Asterisks in each color represent the p values.

G). The percentages of T_em cells of Granzyme B or PD1 expression in total CD45 cells. Blue bar: exhausted T_em (GzmB⁻ PD1⁺); red bar, T_em cells that are GzmB⁺ and PD1⁺; green Bar: functional T_em cells (GzmB⁺PD1⁻) without the feature of exhaustion. Asterisks in each color represent the p values.

H). The percentages of CD8⁺GzmB⁺ T cells distinguished by T-bet and Eomes expression in total CD45 cells. Green bar: effector CD8 (T-bet⁺Eomes⁻); red bar: intermediate state of activated CD8 cells (T-bet⁺Eomes⁺); blue bar: memory CD8 cells (T-bet⁻Eomes⁺); Asterisks in each color represent the p values. n = 4–10.

Tumor samples of Sub T and Liv T were collected from 3 individual experiments. Statistical analysis: one-way ANOVA or Kruskal-Wallis. * p<0.05, **p<0.01.