DNA of mouse teratocarcinoma cells has been analysed as to content of methylated bases by a sensitive method based on two consecutive steps of two-dimensional thin-layer chromatography of radioactively labelled DNA bases. In pluripotent embryonal carcinoma cells (EC), and EC cells cultured under differentiating conditions, as well as teratoma-derived myoblasts and fibroblasts, 5-methylcytosine (5-MC) was the only methylated base found. DNA of the differentiated cell lines (fibroblasts and myoblasts) contained 3.3% and 3.6% 5-MC respectively, while that of embryonal carcinoma cells had 3.8%-5.2%, depending on the cell line. During in vitro differentiation the PCC3/A/1 cell line showed some decrease in percentage of 5-MC (4.2% for EC cells and 3.8% for 30-day cultures). © 1979.