Location of the HSD11B1 promoter methylation assays used in this study. (A) Genome coordinates on chromosome 1, location of individual CpG dinucleotides (dashes) and methylation assay location (black boxes) relative to the 11β-hydroxysteroid dehydrogenase-1 HSD11B1 gene (vertical blue dashes, exons; arrowed lines, introns – according to the UCSC genome browser hg_18 assembly). Arrows denote transcriptional direction. P1 and P2 refer to two independent promoters examined in this study. (B) Higher resolution view of methylation assay 1 (containing a single CpG site adjacent to a conserved putative glucocorticoid response element (GRE)) and assay 2 (containing five CpG sites, three of which were amenable to measurement of methylation levels by Sequenom). (C) Sequence of conserved GRE located within the HSD11B1 P1 region (uppercase letters) relative to the CpG site assayed in this study (red lettering).