Figure 2

5-hydroxymethylcytosine (5hmeC) levels adjacent to the NOS3 TSS correlate with impaired placental perfusion and upregulation of NOS3 mRNA. (A) 5meC (red line) within the NOS3 promoter exhibited a local depression in all patients with minimal variations. In contrast, the amounts of precipitated 5hmeC-enriched DNA (green line) showed a local peak, particularly at the TSS (P4) with significant variations between individuals. In the gene structure cartoon (below), the first 5 NOS3 exons are marked as red boxes and a black arrow indicates the TSS. CpG-enriched regions analysed by qPCR are shown as green boxes (P1 to P6). The blue box marks the location of the 27 nt-ncRNA in intron 5. (B) 5hmeC levels at the TSS (P4), but not at other CpG-rich loci, directly correlate with NOS3 mRNA levels.