Fig. 6

gCpG regulates tumor microenvironment. Tumor-bearing mice were immunized with PBS, CpG + OVA or gCpG on day 7 and 12. On day 19, a frequency of Tregs, b CD8/Treg ratio, percentages of both M1 macrophages (c) and M2 macrophages (d), and prevalence of CD11b⁺Gr1⁺ MDSCs cells (e) in the TME were determined by FACS. f and g IFNγ and TNFα level in supernatants of tumor. h and i Tumor foci of vaccine-treated mice with depletion of macrophages or MDSCs by i.p. injection of 800 µg Clophosome or 250 μg anti-Gr-1 antibody 2 day before vaccination, and the injections were repeated 7 days later. Mice were sacrificed on day 7 post the last immunization and the metastatic nodules were counted. The experiments were performed with 5–7 mice per group. The assays were done in quadruplicates. The data shown are the representative of three experiments. *p < 0.05 and **p < 0.01, ***p < 0.001 and ****p < 0.0001