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. Author manuscript; available in PMC: 2015 Sep 18.
Published in final edited form as: Mol Cell. 2014 Sep 11;55(6):904–915. doi: 10.1016/j.molcel.2014.08.010

Figure 2. A MAFG-Directed Corepressor Complex Mediates MLH1 Transcriptional Silencing.

Figure 2

(A) qRT-PCR analysis monitoring MLH1 expression in RKO cells expressing a NS, BACH1, BACH2 or NFE2L1 shRNA.

(B) qRT-PCR analysis monitoring MLH1 expression in RKO cells expressing a NS, DNMT1, DNMT3A or DNMT3B shRNA.

(C) Bisulfite sequencing analysis of the MLH1 promoter in RKO cells expressing a NS, MAFG, BACH1, CHD8 or DNMT3B shRNA. Decreased promoter methylation in the presence of 5-AZA is shown as a control.

(D) ChIP analysis monitoring binding of MAFG, BACH1, CHD8 and DNMT3B to the MLH1 promoter or, as a control, an irrelevant DNA region (negative control [NC] DNA) in RKO cells. The results were normalized to that obtained with IgG, which was set to 1.

(E) ChIP analysis monitoring binding of MAFG, BACH1, CHD8 and DNMT3B to the MLH1 promoter in RKO cells expressing a NS, MAFG, BACH1, CHD8 or DNMT3B shRNA. Data are represented as mean ± SD. *P≤0.05, **P≤0.01.

(F) Co-immunoprecipitation analysis. RKO cell extracts were immunoprecipitated with a MAFG, BACH1, CHD8, DNMT3B or control (IgG) antibody, and the immunoprecipitate was analyzed for MAFG, BACH1, CHD8 or DNMT3B by immunoblotting. See also Figure S2.