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. 2004 Mar 23;90(8):1606–1611. doi: 10.1038/sj.bjc.6601716

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Copyright © 2004 Cancer Research UK

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Methylation analysis of the GPC3 promoter in nontumoural samples. (A) CpG dinucleotides positions in the GPC3 promoter region. The methylation status of 11 of these CpG sites was determined either by the PCR-based methylation assay (#) or by the Southern blot-based methylation assay (+) using methyl-sensitive restriction endonucleases HpaII (H), SacII (S), EagI (E), and BssHII (B). HpaII contains one CpG site, whereas SacII, EagI and BssHII contain two CpG sites each. The distal and proximal sites were amplified in distinct PCRs. (B and C) Representative results of the methylation analysis in normal peripheral blood and placental DNA samples obtained by the PCR-based (B) and Southern blot-based (C) methylation assays. See Table 1 for details concerning the samples. (D) PCR-based methylation assay performed on DNA samples from two individuals (AC-1 and DC) affected by the Turner syndrome. Digestions: (B and D) H, HpaII; M, MspI; U, undigested; (C) H, HindIII; B, BssHII; S, SacII and E, EagI. GPC3+, expression of GPC3; GPC3−, no expression of GPC3.