Fig. 4

DIM treatment enhanced antitumor immune responses of anti-PD-1 in 4T1 tumor-bearing mice. A Representative images of tumors, B tumor volume and C tumor weight were shown after 4T1 tumor-bearing mice were treated with 10 mg/kg DIM three times a week for 2 weeks from day 8 after tumor cell inoculation and were intraperitoneally injected with anti-PD-1 mAb (0.25 mg/mouse) or isotype control antibody every two days from day 11 to 19 after tumor cell inoculation (n = 8). D The percentage of total MDSCs and their subsets from bone marrow, blood, spleen and tumor was analyzed by flow cytometry. E Relative mRNA levels of Arg1 and iNOS in MDSCs from spleen of 4T1 tumor-bearing mice after co-treated with DIM and anti-PD-1. F The percentage of CD4⁺ and CD8⁺ T cells from blood, spleen and tumor in 4T1 tumor-bearing mice after co-treated with DIM and anti-PD-1. G IFN-γ level in tumor was measured by ELISA. Data are representative results from three independent experiments and the results are expressed as the mean ± SEM. *p < 0.05, **p < 0.01 and ***p < 0.001. T-MDSC: total MDSCs; M-MDSC: monocytic MDSC; G-MDSC: granulocytic MDSC