This study aimed to examine whether the relationship between mothers’ DNA methylation profiles and offspring's depression is modulated by negative parenting. The participants were 35 African-American mother–offspring dyads. Young adult offspring (19 females; age = 17–29.5 years) were assessed on depressive symptoms, and mothers (36–51 years) were assessed on negative parenting. Methyl-binding domain (MBD) sequencing was used to assay genome-wide DNA methylation in peripheral T lymphocytes. Controlling for the effect of offspring's DNA methylation, mothers' DNA methylation was positively associated with offspring's depression. Hypermethylation of a subset of the epigenetic markers was associated with increased negative parenting. Negative parenting was positively correlated with offspring's depression as well, suggesting that negative parenting may be a modulator between the mother's epigenome and offspring's depression. This study is one of the first investigations of the modulating role of parenting behavior in associations between the mother's epigenome and offspring's depression.