Fig. 4

Ex vivo analysis of tumor samples from the 3LL syngeneic mouse model. (A) Left panel, representative FACS plots and gating strategy. Right panel, bar graphs of tumor-infiltrated T cells analyzed using FACS. Percentages of CD4⁺ T cells (defined as Live/Dead⁻CD45⁺CD3⁺CD4⁺) and CD8⁺ T cells (defined as Live/Dead⁻CD45⁺CD3⁺CD8⁺) in total live cells (defined as Live/Dead⁻) (n = 6/group). The representative flow images and gating strategy of 4 different groups are shown in Supplementary Fig. S7. (B) The representative images of the IHC double staining using anti-PD-1 and CD8 antibodies are shown, Red: PD-1; Brown: CD8. (left panel, scale bar: 50 μm). The average percentages of the double-positive staining of PD-1⁺CD8⁺ T cells were analyzed (right panel, n = 10/group). (C) The representative images of the IHC staining using anti-CD31 antibody are shown (left panel, scale bar: 50 μm). The CD31 IHC data were quantified (right panel, n = 6 for each group). The error bars represent SEM; *, P < 0.05