Fig. 2

DNA methylation at the Slc17a6 promoter. a Schematic representation of the Slc17a6 gene. Solid boxes represent the 5′ and 3′UTRs and open boxes represent exons. A region spanning −255 to +90 bp relative to the transcriptional start site (TSS) is drawn below. The region (BS1, −144 to −20 bp) and CpG sites (vertical arrows, CpGs 1–9) examined by clonal bisulphite sequencing are indicated, as is the region used in in vitro reporter assays (CpGs 1–10, −144 bp to +68 bp). Four different CpG methylation patterns were generated for reporter assays using three bacterial methyltransferases (MTs), M.SssI MT, HhaI MT and HpaII MT either alone or in combination. Filled and open circles represent methylated and unmethylated CpGs, respectively. b DNA methylation in the adult hippocampus (P87 and 120). Each line of nine CpGs represents the DNA methylation state of one allele in one cell. Clones from controls (total n = 42 from 16 mice) as well as basal (total n = 33 from 16 mice) and high (total n = 34 from 17 mice) Slc17a6 expressors in the ethanol-exposed (EtOH) group are shown. A graph of % DNA methylation per clone is also shown. c The impact of Slc17a6 promoter methylation (CpGs 1–10, −144 to +68 bp) on luciferase activity in vitro. The results of three independent experiments in CAD cells are shown. Data are presented as mean and standard deviation. **P < 0.01 (t test, two-tailed)