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Fig. 1 | Genes & Nutrition

Fig. 1

From: γ-Glutamyl hydrolase modulation significantly influences global and gene-specific DNA methylation and gene expression in human colon and breast cancer cells

Fig. 1

Effects of GGH modulation on global DNA methylation and DNMT activity in HCT116 and MDA-MB-435 cells. In both cell lines, GGH overexpression was associated with significantly lower global DNA methylation (HCT116, a; MDA-MB-435, c) and lower DNMT activity (HCT116, e; MDA-MB-435, g) compared with controls expressing endogenous GGH, whereas GGH inhibition showed significantly higher global DNA methylation (HCT116, b; MDA-MB-435, d) and higher DNMT activity (HCT116, f; MDA-MB-435, h) compared with controls expressing endogenous GGH. The in vitro methyl acceptance assay for global DNA methylation measurement produces an inverse relationship between the endogenous DNA methylation status and exogenous [3H-methyl] incorporation into DNA. All analyses were performed in quadruplicate and repeated using two independent cell lysates (ad). The assay for DNMT activity produces a positive relationship between the endogenous enzyme activity and exogenous [3H-methyl] incorporation into DNA. Each reaction was performed in triplicate, and the assay was repeated three times (eh). Control-S, cells expressing endogenous GGH; Sense, cells transfected with the sense GGH cDNA; Control-si, cells expressing endogenous GGH; siRNA, cells transfected with the GGH-targeted siRNA. *P < 0.05; **P < 0.01; ***P < 0.001 compared with corresponding control by the Student’s t-test. Values are mean ± SD

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