Figure 4. Expression of Suz12 and Tet genes correlated with the presence of methylcytosine (5mC) and hydroxymethylcytosine (5hmC) at the Avp locus during hypothalamic-like differentiation.

A, Suz12 gene expression rapidly decreased following 4 days of Lif withdrawal. B, Similarly, Tet1 and Tet2, but not Tet3, gene expression rapidly declined following 4 days of Lif withdrawal. C, MeDIP showed high hydroxymethylation (5hmC) at the Avp gene body and downstream enhancer which strongly decreased upon Lif-withdrawal. Application of cyclopamine evoked a further decline at the enhancer region. D, Conversely, methylation (5mC) at the Avp enhancer, but not at the gene body, increased as evidenced by MeDIP. Bars represent standard deviations from the mean (sem), *, P<0.05; from3 independent experiments performed in duplicate.