Figure 3.MAGEA11 promoter methylation and expression in prostate cancer and EOC. (A) Sodium bisulfite clonal sequencing of the MAGEA11 promoter region was performed on the indicated cell lines. The transcriptional start site (TSS), as determined by RLM-RACE, is indicated by the right broken arrow, and the coordinates of the analyzed region are shown in the upper left panel. Filled and open circles indicate methylated and unmethylated CpG sites, respectively, and each row represents one sequenced allele. The red box indicates three TSS-adjacent CpG sites. (B) Summary of MAGEA11 bisulfite sequencing and mRNA expression data in prostate cell lines. Methylation percentages for the entire MAGEA11 5′ CpG island (CGI) or for the three TSS-resident CpGs were calculated from (A), and MAGEA11 expression was determined by RT-qPCR. (C) MAGEA11 promoter methylation and mRNA expression in one normal ovary (NO) and four EOC samples were determined using bisulfite clonal sequencing (> 10 alleles), and Affymetrix microarray, respectively. (D) MAGEA11 mRNA expression indirectly correlates with MAGEA11 TSS methylation in EOC. MAGEA11 expression and methylation of three TSS-resident CpG sites in 16 EOC samples was determined by RT-qPCR and bisulfite pyrosequencing, respectively. Five samples that did not express measurable MAGEA11 are plotted on the x-axis. Spearman test results are shown.