Fig. 5

RES treatment modulates the tumor immune microenvironment in the ID8 tumor model. a Flow cytometry gating and histogram analysis of matured dendritic cells (DCs) in the tumor tissues at the end of treatment. The matured DCs were denoted as CD80⁺CD86⁺ populations (gate in CD45⁺CD11b⁺CD11c⁺ cell population). Data represent means ± SD, n = 8, **p < 0.01, ***p < 0.001, versus Control group; (b) Flow cytometry gating and histogram analysis of cytotoxic T cells (CD8⁺ T cells) in the CD45⁺ tumor infiltrating immune cells. Data represent means ± SD., n = 8, **p < 0.01, ***p < 0.001, versus Control group; ELISA results of TGF-β (c), IL12p70 (d) and IFN-γ (e) production in the tumors from mice receiving PBS or RES treatments. Data represent means ± SD. n = 8, *p < 0.05, **p < 0.01 ***p < 0.001, versus Control group. f Antitumor effect of combination of RES and aCD8 or aPD-1. To demonstrate the critical role of cytotoxic CD8⁺ T cells in antitumor immunity, anti-CD8 monoclonal antibody was i.p. injected in treatment group 3 days prior to the first drug administration and repeated every 3 days until the termination of the study, and anti-PD1 monoclonal antibody was i.p. injected in treatment group 7 days post first injection, and repeat 3 times. The injection dosages of aCD8 and aPD-1 were 200 μg/mouse and 100 μg/mouse, respectively. Data represent means ± SD. n = 8, **p < 0.01