Figure 6

Identification of a hypermethylated region within the IG-DMR of GCTSCs. Cellular DNA was extracted from GCTSCs (n = 8) and MSCs (n = 8) and DNA fragments covering the Meg3-DMR (44 CpGs) and the IG-DMR (31 CpGs) were amplified by PCR, bisulfite treated, cloned into pCR4-TOPO vector and sequenced. (A, B) Calculated methylation frequencies of all analyzed CpGs within the Meg3-DMR and the IG-DMR of 10 individual clones derived from one GCTSC and one MSC cell line. (C, D) Calculated methylation frequencies within the Meg3-DMR and the IG-DMR of eight different GCTSC and MSC cell lines. (E) Methylation analysis restricted to the first 13 CpGs analyzed within the IG-DMR. Data are presented as mean ± SD. (*p < 0.05 determined by Mann–Whitney-U test).