Figure 2.

Schematic representation of the H19 gene. The position of H19 ICR, number of CpG nucleotides analyzed for methylation status and position of primer set used for expression analysis are indicated. For methylation analysis of H19 ICR, two regions were studied: ICR^{CTCF 1–2} (574 bp) and ICR^{CTCF 3–4} (660 bp). The positions of the CpGs analyzed in each sequence are shown by lollipops (19 CpGs in ICR^{CTCF 1–2} and 23 CpGs in ICR^{CTCF 3–4}, respectively). The four CTCF binding sites are depicted as grey boxes within the ICR^{CTCF 1–2} and ICR^{CTCF 3–4}. For proximal part of H19 promoter (white box with PP), 8 CpGs were analyzed. For expression analysis, real time quantitative RT-PCR was performed using the Taqman technology with two primers (black arrows) chosen to encompass an intron (wedge indicates the exon-exon splice junctions) and a universal probe (white box).