One-carbon metabolic genes and 5mC levels (%) in ECM-attached and -detached conditions. (a) The mRNA expression levels of one-carbon metabolic genes in the attached (AT) methionine cycles and 6 days detached (6D_DT) conditions in the HeLa cell lines. (b) The mRNA expression levels of one-carbon metabolic genes in folate cycles in attached (AT) and 6 days detached (6D_DT) conditions in the HeLa cell lines. (c) The mRNA expression levels of one-carbon metabolic genes in methionine cycles in attached (AT) and 6 days detached (6D_DT) conditions in the MCF-7 cell lines. (d) The mRNA expression levels of one-carbon metabolic genes in folate cycles in attached (AT) and 6 days detached (6D_DT) conditions in the MCF-7 cell lines. SHMT1, serine hydroxymethyltransferase 1; MTHFR, methylenetetrahydrofolate reductase. (e,f). HeLa and MCF-7 cell lines showed increased 5mC levels in the 6 days (6D_DT) ECM-detached condition compared to the attached (AT) condition. The data are presented as mean ± SEM; * p < 0.05, ** p < 0.01, *** p< 0.001, and **** p < 0.0001.