Figure 3

TSDR-DMR, expression levels of FOXP3 mRNA and protein in tissue samples. (A) Significantly higher TSDR-DMR in tumor samples versus normal samples, as investigated by MS-qPCR (p = 0.000, n = 130). (B) Significantly higher FOXP3 mRNA expression in samples of tumor tissues versus samples of normal tissues, as detected by real-time RT-PCR (p = 0.003, n = 21). The Wilcoxon matched pairs test was applied to both (A) and (B) because the data were not normally distributed. A two-tailed p value ≤0.05 was considered statistically significant. (C) Representative results showing higher FOXP3 mRNA expression in tumor tissues versus normal tissues, as detected by agarose gel electrophoresis (p = 0.009, n = 5). RT-PCR products of target (FOXP3) and internal control (β-actin) gene were loaded in two rows of parallel lanes and electrophoreses were performed simultaneously. DL500 was served as a DNA marker. (D) Representative result showing increased FOXP3 protein synthesis in tumor tissues versus normal tissues, as detected by Western blotting (p = 0.039, n = 4). For both (C) and (D), ImageJ software was used to perform quantitative analysis of the densities of band, and paired t tests were applied. A two-tailed p value ≤0.05 was considered statistically significant.