Promoter hypermethylation and lack of PRDX2 expression in Hodgkin and Reed-Sternberg (HRS) cells. (A) Localization of the 27K BeadArray (Illumina) array tags cg03860466 and cg00155609 (indicated by arrows) with respect to the PRDX2 TSS. (B) PRDX2 mRNA expression (Affymetrix U95 microarray, tag 39729_at) in cHL cell lines, normal B-cell populations (10 × GCB cells, 5 × naive B cells, 5 × memory B cells), 15 diffuse large B-cell lymphomas (DLBCLs), 11 Burkitt lymphomas (BLs), 6 follicular lymphomas (FLs), 11 B-cell chronic lymphocytic leukemias (CLLs), and 5 large cell lymphomas (LCLs). Mean PRDX2 promoter methylation level based on 27K BeadArray (Illumina) array (tags: cg03860466, cg00155609) in the cHL cell lines is given in parentheses. Methylation and expression data were taken from Ammerpohl et al2 and Küppers et al,3 respectively. (C) Bisulfite pyrosequencing of PRDX2 promoter region in cHL cell lines. Bars represent the cytosine guanine dinucleotide (CpG) sites analyzed within the sequenced region (genomic position of the CpG sites: black bar, chr19:12912528; gray bar, chr19:12912542). No bar indicates a methylation level of 0%. (D) Bisulfite pyrosequencing of PRDX2 promoter region in microdissected HRS cells (nodular sclerosis, cases 1, 2, 4, 5; mixed cellularity, 3) and nontumoral bystander cells (Lymph) (in duplicate) (genomic position of the CpG sites: black bar, chr19:12912528; gray bar, chr19:12912542). No bar indicates a methylation level of 0%. M, methylated DNA control (Millipore); POOL, whole blood DNAs, pooled; U, unmethylated DNA control (whole genome amplified DNA).
