We observed globally altered DNA methylation upon decitabine exposure and withdrawal. Since DNA methylation is a major epigenetic mechanism that is involved in modulating gene expression and chromatin architecture [22], these observed changes in 5mC levels may possibly lead to altered chromatin structure and genome-wide changes in gene expression. Furthermore, the presented findings highlight that exposure to drugs that disturb the epigenetic marks during differentiation of brain cells may lead to aberrant DNA methylation profiles. Our observations at D8 indicate that, even after the disturbance factor is withdrawn from the system, an epigenetic memory for this disturbance may be associated throughout cellular differentiation of brain cells. Thus, our findings highlight the biological importance of maintaining proper regulation of epigenetic factors/modifications during brain development with a clear focus on DNA methylation and MeCP2.