Fig. 7

Immunohistochemical staining assays to measure the infiltration of T cells in EMT-6 tumors. a–d The presentative images of tumor-infiltrating CD3⁺ cells in the tumor periphery and the tumor center. Scale bars, 250 μm or 50 μm. e The quantitative analyses for the number of tumor-infiltrating CD3⁺ cells, and the proportion of CD3⁺ area was used. f The quantitative analysis for the infiltration depth. The infiltration depth of CD3⁺ cells was calculated by the mean nearest distance of all CD3⁺ cells to the tumor border. The mean nearest distance was scaled by the distance between the corresponding tumor border to tumor center. g, h The presentative images of tumor-infiltrating CD4⁺ and CD8⁺ cells in the tumor periphery and the tumor center. Scale bars, 100 μm. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 denote the significant difference relative to YM101 treatment. α-TGF-β: anti-TGF-β, α-PD-L1: anti-PD-L1