A) Dissection and isolation of myenteric plexus from P0 wild-type and Ezh2 null mice for qRT-PCR and ChIP assays. B) qRT-PCR of Pax3, Zic1 and Sox10 in myenteric plexus. In the Ezh2 null myenteric plexus, Pax3 was upregulated 20–40 fold (n = 3, p = 0.0010), Sox10 was upregulated 5–30 fold (n = 3, p = 0.0263), and Zic1 was upregulated 40–100 fold (n = 3, p = 0.0071) compared to its wild type littermates. C) H3K27me3 ChIP-qPCR in Ezh2 mutant (in orange) and wild-type littermate (in grey) P0 gut tissues. The amount of precipitated DNA in promoter regions of NCC developmental genes and HoxA9 are presented as a relative value (%) to that of the input DNA normalized over a negative locus (beta actin) (y-axis). Student’s t-test (two-tailed) value shows significant difference in H3K27me3 enrichment between Ezh2 null (n = 6) and wild-type (n = 8) littermates (p<0.01).