MOHANDAS et al. (1981) have proposed that mammalian X chromosome inactivation involves segmental methylation of cytosine residues in the DNA at multiple sites along the X chromosome. Using antibodies specific for 5-methylcytosine, we have found no detectable difference in the extent of methylation of the DNA in the two X chromosomes of owl monkey or human females. Thus inactivation (facultative heterochromatization) of the X chromosome is not due to, or associated with, intense DNA methylation comparable to that seen in most constitutive heterochromatin.