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Fig. 4 | Clinical Epigenetics

Fig. 4

From: The absence of AhR in CD4+ T cells in patients with acute graft-versus-host disease may be related to insufficient CTCF expression

Fig. 4

CTCF recruits TET2 and EP300 to interact with the AhR promoter region. A Co-immunoprecipitation using anti-CTCF antibodies in Jurkat cells after CTCF and TET2 expression, for the detection of CTCF binding and TET2 by Western blot. B, C ChIP-qPCR analysis of the enrichment levels of TET2 (B) and EP300 C in the AhR promoter in chromatin fractions extracted from CD4+ T cells from patients with aGVHD (n = 10) and the non-aGVHD group (n = 10). Results are relative to input DNA prepared from untreated chromatin. D Normal CD4+ T cells were divided into three groups and transfected with control pRS and pCMV6 plasmids or CTCF silencing (pRS-CTCF) and TET2 and EP300 expression (pCMV6-TET2 and pCMV6-EP300) plasmids, or pRS and pCMV6-TET2 and pCMV6-EP300, respectively. CTCF, TET2 and EP300 protein levels were detected by Western blot. E ChIP-qPCR analysis of TET2 and EP300 enrichment levels in the AhR promoter. Results are relative to input DNA prepared from untreated chromatin. Data represent the means of three independent experiments. F aGVHD CD4+ T cells were divided into two groups and transfected with control pCMV6 and CTCF expression (pCMV6-CTCF) plasmids. CTCF protein levels were detected by Western blot. G ChIP-qPCR analysis of TET2 and EP300 enrichment levels in the AhR promoter. Results are relative to input DNA prepared from untreated chromatin. Data represent the means of three independent experiments (*P < 0.05, **P < 0.01)

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