Fig. 7

Etoposide enhances the therapeutic efficacy of Tim-3 blockade therapy. a Schematic diagram illustrating the treatment protocol of Tim-3 antibody (Tim-3 Ab) and/or etoposide (ETO) in mice. At the endpoint, tumor cells and tumor-infiltrating lymphocytes (TIL) were isolated for analysis. b Tumor growth of 4T1 cells in BALB/c mice treated with Tim-3 antibody and/or etoposide. Tumor size was measured at the indicated time points and tumor weight was measured at the endpoint (n = 6 mice per group). c Efficacy of indicated treatments on the CSC (CD44⁺CD24⁺ALDH1⁺ populations) frequency of 4T1 tumors. d Flow cytometric analysis analyzing the effect of indicated treatments on the expression of E-cadherin (E-Cad, epithelial marker) and N-cadherin (N-Cad, mesenchymal marker) in the entire cancer cell population of 4T1 tumors. e qRT-PCR analysis of the influence of indicated treatments on STT3 isoforms expression in the entire cancer cell population of 4T1 tumors. f Effect of indicated treatments on PD-L1 expression in the CSC and non-CSC populations of 4T1 tumors. g Intracellular cytokine staining of CD8⁺ IFNγ⁺ cells in the CD3⁺ T cell populations from isolated tumor-infiltrating lymphocytes to analyze the impacts of indicated treatments on tumor-infiltrating cytotoxic T cell activity. Error bars represent s.d. (n = 6). *P < 0.05; ns: non-significant, Student’s t-test. See also Supplementary Fig. 7