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. 2007 Oct 8;27(23):8164–8177. doi: 10.1128/MCB.00555-07

FIG. 5.

FIG. 5.

Differential Wnt inducibility correlates with distinct epigenetic states of the corresponding target genes. (A) Analysis of DNA methylation patterns at the Axin2, Cdx1, and T/Bra promoters. Genomic DNA was isolated from C17.2, C2C12, or E14 cells after treatment for 24 h with 25 μM (C17.2, C2C12) or 5 μM (E14) SB-216763 or with DMSO as the solvent control. Bisulfite-treated DNA fragments of the promoter regions were PCR amplified with primer pairs shown in Fig. 1, subcloned, and analyzed by sequencing. Each line represents the results from a single sequence and each circle denotes a CpG dinucleotide (filled circles, methylated CpGs; open circles, nonmethylated CpGs). (B) ChIP analysis for acetylated histone H3 in C17.2, C2C12, or E14 cells. Formaldehyde-fixed chromatin was prepared after treatment with 25 μM (C17.2, C2C12) or 5 μM (E14) SB-216763 or with DMSO as the solvent control for 24 h. Chromatin was precipitated with 2 μg of an anti-acetylated histone H3 (α-H3ac) antibody or equal amounts of a nonspecific rabbit isotype control. ChIP DNA was analyzed by quantitative real-time PCR with primer pairs within the Axin2, Cdx1, and T/Bra promoter regions. Nucleotide positions of the amplicons relative to the transcriptional start site are indicated (Fig. 1). Bars represent relative enrichment compared to that of the nonspecific isotype control (relative enrichment of 1). Data given are the average and SEM from at least three experiments.