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Fig. 3 | Clinical Epigenetics

Fig. 3

From: DNA methylation instability by BRAF-mediated TET silencing and lifestyle-exposure divides colon cancer pathways

Fig. 3

TET1 is suppressed in BRAFV600E cancer cell lines. a Unsupervised hierarchical clustering using DNA methylation levels of CCN-CpGs (1312) and CIMP-CpGs (1800) in colon cancer cell lines measured by HM27K/HM450K. Shown are the CpGs that were present on both arrays. Heatmap of the β-values (upper) with boxplot of resultant β-values (lower). Note the increase DNA methylation in cell lines with BRAFV600E. bTET1, TET2, and hMLH1 mRNA expression in colon cancer and in normal colon epithelial cell lines (CCD841CoN). Error bars denote SD (n = 3). Primers were designed to measure both isoforms of TETs. c β-values of TET1 and TET2 CpGs measured by HM27K/HM450K. dTET1, TET2, and hMLH1 mRNA expression following treatment with 0.1 μM 5-azacytidine (Aza) or dimethyl sulfoxide (DMSO), presented as relative expression compared to DMSO. P values were calculated with Welch two sample t-test. Error bars denote SD (n = 3). e Western blot analysis of TET1 and TET2. Indicated with asterisks are the full-length (TET1FL) and alternative (TET1ALT) TET1 and two isoform of TET2 (a, b). Signal was quantified by image studio software and shown as ratio to GAPDH for TET1 (TET1FL+ALT) for TET2 (TET2a and 2b). Shown are the representative blot from four (for TET1) or two (for TET2) independent experiments. f Levels of 5hmC measured by dot blot analysis with methylene blue staining (DNA) as loading control

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