Aberrant methylation at the FZD9 locus is a predictor of prognosis in MDS/AML. (A) FZD9 hypermethylation is associated with decreased 12-month survival. (B) FZD9 methylation levels in individual patients classified by disease group. Each dot represents the methylation β-value of the FZD9 CpG site in an individual patient. The average methylation β-value of this CpG site for each patient group is indicated by the horizontal line. (C) FZD9 methylation is inversely correlated to FZD9 transcript expression. represents the degree of FZD9 methylation in the sample; ■, the FZD9 expression levels determined by semiquantitative RT-PCR, defined as fold change compared with the control marked with an asterisk. (D) FZD9 CpG methylation is not part of a wider hypermethylation at its chromosome 7 locus. The height of the vertical bars represents the frequency of aberrant hypermethylation of the CpG sites designated along the horizontal line. The space between bars marked with 0 indicates analyzed CpG sites that were not hypermethylated in any patient. (E) Regions of chromosome deletion and UPD, detected by SNP-A, that involves the FZD9 locus. FZD9 is designated by the vertical red bar. Red lines depict single SNP signal intensity, whereas green lines present an average value of SNP signal intensity. The horizontal purple bar represents areas of chromosome deletion, whereas the horizontal blue bar represents areas of loss of heterozygosity through UPD. (F) Chromosomal deletion or duplication of an FZD9 allele, combined with aberrant methylation of the remaining allele, is associated with a worse prognosis than either abnormality alone.