Fig. 3

DAC downregulated T-cell exhaustion and upregulated T-cell infiltration by demethylating Runx3 and promoting Runx3 expression. a Workflow of the experimental design and analysis of the tumor growth curve using the MC38 mouse model treated with DAC, anti-PD-1 or DAC/anti-PD-1(n= ). b Tumor growth curve of mice treated with DAC, anti-PD-1 or anti-PD-1/DAC. Upperpanel: average tumor growth curves;(two-tailed unpaired t tests, *P <0.05, **P<0.01, ***P<0.001) c The proportions of GranB+, perforin+, TNF-α+, IFN-γ+, Ki67+ and CD8+ T cells were analyzed by flow cytometry. Samples were taken from the blood, spleen, tumor, or lymphocytes of MC38 mice treated with DAC, anti-PD1 or anti-PD-1/DAC as indicated. (n=5, two-tailed unpaired t tests, *P <0.05,**P <0.01***P < 0.001). d Leftpanel: The proportion of Runx3+CD8+ T cells in each group was analyzed by flow cytometry (n=5, two-tailed unpaired t tests, **P<0.01). Right panel: DNA methylation level change on Runx3 promoter in T cells treated with DAC, anti-PD-1 or antiPD-1/DAC. Y axis: Mehylation level of Runx3 (%); X axis: sampes of mice treated with DAC, anti-PD-1 or DAC/anti-PD-1. Triplicate samples were applied for each experiment and the median was shown as horizontal line within the box plots. (p<0.05)