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Figure 4 | Epigenetics & Chromatin

Figure 4

From: Menin and RNF20 recruitment is associated with dynamic histone modifications that regulate signal transducer and activator of transcription 1 (STAT1)-activated transcription of the interferon regulatory factor 1 gene (IRF1)

Figure 4

Dynamic H3 lysine methylation at the activated interferon regulatory factor 1 gene ( IRF1 ) and IRF1 transcription are altered by 5'-deoxy-5'-methyl-thioadenosine (MTA) treatment. 2fTGH cells treated with MTA (1.5 mM) or dimethylsulfoxide (DMSO) for 24 h followed by treatment with interferon (IFN)γ for 30 min. (a-c) Chromatin immunoprecipitation (ChIP) using the indicated antibodies. Data were normalized to Pan H3 cycle threshold (Ct) values to account for a non-specific carrier (DMSO) effect and reported as percentage of input. (d, e) Reverse transcription quantitative PCR (RT Q-PCR) was performed after the times shown to quantitate IRF1 mRNA and pre-mRNA expression relative to glyceraldehyde 3-phosphate dehydrogenase gene (GAPDH) expression and presented as fold change upon induction. Error bars represent standard error (n = 3). **P = 0.01, *P = 0.05. (f) Arrows indicate locations of Q-PCR primers designed to amplify either exonic (d) or intronic (e) regions of the IRF1 gene. (g, h) RNA polymerase II (Pol II) and signal transducer and activator of transcription 1 (STAT1) ChIP of 2fTGH cells.

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