Spry2 is DNA-hypermethylated, and its expression is repressed in TCL1-tg B-cell lymphomas. (A) Spry2 RLGS locus (arrow) is shown for WT mouse spleen and 3 TCL1-tg B- cell tumors. Comparisons to the WT spot intensity scored BLL3 as unmethylated/partially methylated (> 50% of WT) and BLL1a and MZL1 as DNA hypermethylated (< 50% of WT) for the surveyed NotI site.^14,40  (B) GBS of a CpG-rich region located between −728 and −484 in the mouse Spry2 promoter. Each row represents the sequence of an individual clone of WT spleen, MZL2, or BLL1a cells. ○, represents unmethylated CpG sites; ●, methylated sites; squares, 2 CpG loci within the Not1 site. (C) Spry2 expression in WT or TCL1-tg B-cell lymphomas was determined by quantitative PCR. Average Spry2 expression for each group (WT, hypermethylated, and unmethylated/partially methylated lymphomas) is shown by a dashed line. Spry2 expression was normalized to 36b4 (Rplp0) expression. DNA methylation status for WT or TCL1-tg B-cell lymphomas was also assessed by MSP. The top and bottom bands correspond to PCR products amplified by primers specific for methylated (M) or unmethylated (U) CpG sites after bisulfite conversion.