Figure 1

a. Schematic view of the generation of a Not I MSDK library. The black lollipops indicate methylated Not I sites that will not be digested by the enzyme. 1b. The number of virtual tags (lined bars) per chromosome compared to the number of experimental tags (black bars) shows an unbiased sampling of the mouse genome. On average > 80% of experimental tags could be matched to a virtual tag after removal of all singletons from the pool of Not I MSDK genomic tags. 1c. A log10 scale scatter plot comparison of genomic tag counts between the HMD (y-axis) and the LMD (x-axis) MSDK libraries. Significant differentially methylated genomic tags with a P-value = 0.05 with higher counts in HMD are indicated by green triangles and those with higher counts in LMD are indicated by black diamonds. Points colored in grey are considered non-significant (P-value > 0.05). 1d. Snapshot UCSC genome Browser displaying the location of the genomic tag found to be differentially methylated and associated with Tmem151a on chromosome 19 (Table 2). The Not I restriction sites are also indicated. MSP was performed and the PCR products were sub cloned into a TOPO TA sequencing vector. Up to 12 independent clones with insert were selected for sequencing. The Sequence analysis of bisulfate modified genomic DNA from 2 HMD lung tissue samples revealed a methylated Not I site in the proximity of the gene encoding hypothetical protein Tmem151a in 2 out of 12 sequenced clones.