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. 2010 Oct 20;96(1):E199–E207. doi: 10.1210/jc.2010-1647

Figure 4.

Figure 4

Fine-scale determination of the methylation status of CGB5 promoter region in placenta and blood using bisulfite sequencing. A, Localization of analyzed CpG sites (vertical lines) in the upstream and transcribed region of CGB5. Positions of primers (Supplemental Table 1) for the amplification and clonal sequencing of methylated (V, S) and unmethylated (Z, U) promoter sequence from bisulfite-modified DNA are indicated. One differentially methylated CpG site is located within the Sp1 binding site, critical in the basal HCGβ expression (19). B, The methylation status of 13 CpG sites within the analyzed region of the CGB5 locus in parental blood DNA (almost complete methylation), placental samples from term pregnancies (NP 44, NP11; almost complete unmethylation) and from placentas with monoallelic expression of the maternal CGB5 allele (RM10, RM9, ETP26; hemimethylation). Four to 10 clones were sequenced per each sample, and the methylation-specific primer combination (UNMET, MET) showed consistent methylation patterns. Filled circle, methylated CpG site; open circle, unmethylated CpG site. Sp1, Selective promoter factor 1.