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. 2010 Jan 5;102(2):419–427. doi: 10.1038/sj.bjc.6605505

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Copyright © 2010 Cancer Research UK

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Validation of three hypermethylated candidate genes in primary TGCT samples. (A) Hypermethylation of the promoters of APOLD1, PCDH10 and RGAG1 in NT2 cells. Hypermethylation of these genes is confirmed by genomic bisulphite sequencing. (B) Downregulated expression of APOLD, PCDH10 and RGAG1 in primary TGCT. NT: normal testis (n=8); EC: embryonal carcinoma (n=9); SE: seminoma (n=8); YST (n=1): yolk-sac tumour. The mean value of each group is represented by the horizontal bar. ^*P<0.005; ^**P<0.05; ^***P<0.001 by two-tailed Student's t-test. (C) Promoter hypermethylation of APOLD1 in primary TGCT. MSP is performed to compare the relative methylation of each patient. In all, 71% of TGCT patients are partially methylated (n=17), whereas none of the subjects with a normal testis (n=6) are methylated. One case each from the tumour group (1289) and normal group (A419) is selected and confirmed by bisulphite sequencing. U: unmethylated; M: methylated. (D) Expression of APOLD1 in other tumours. RNA samples of each tumour and normal adjacent tissue were isolated from a single individual. Error bars indicate s.e.m. of triplicate experiments.