STAT5 and TET2 Cooperate to Regulate FOXP3-TSDR Demethylation in CD4⁺ T Cells of Patients with Colorectal Cancer

Figure 3

TET2 binds to the FOXP3-TSDR and regulates its DNA methylation via STAT5. (a-b) ChIP-qPCR analysis of the enrichment of STAT5 (a) and TET2 (b) in distal promoter, CNS1, and TSDR of FOXP3 in CD4⁺ T cells from colon tumor tissues () and normal colonic tissues (). (c, left) Representative Western blot results for STAT5, TET2, and FOXP3 in CD4⁺ T cells from colon tumor tissues after interference with STAT5 expression. GAPDH was used for normalization. (c, right) Quantitative analysis of the band intensities for STAT5, TET2, and FOXP3 protein levels normalized by GAPDH. The same experiments were repeated 3 times. (d) ChIP-qPCR analysis of the enrichment of TET2 in the FOXP3-TSDR in CD4⁺ T cells from colon tumor tissues after interference with STAT5 expression. The same experiments were repeated 3 times. (e) DNA methylation of FOXP3-TSDR in CD4⁺ T cells from colon tumor tissues after interference with STAT5 expression. The same experiments were repeated 3 times. (f, left) Representative Western blot results for STAT5, TET2, and FOXP3 in CD4⁺ T cells from normal human donors after overexpression of TET2 and inhibition of STAT5. GAPDH was used for normalization. (f, right) Quantitative analysis of the band intensities for STAT5, TET2, and FOXP3 protein levels normalized by GAPDH. The same experiments were repeated 3 times. (g) ChIP-qPCR analysis of the enrichment of TET2 in the FOXP3-TSDR in CD4⁺ T cells from normal human donors after overexpression of TET2 and inhibition of STAT5. The same experiments were repeated 3 times. (h) DNA methylation of FOXP3-TSDR in CD4⁺ T cells from normal human donors after overexpression of TET2 and inhibition of STAT5. The same experiments were repeated 3 times (, ).

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