Figure 4.

CpG methylation of the candidate differentially methylated region (DMR) for the new candidate imprinted gene AXL. (A) Genomic organization of the human AXL promoter region showing the relative location of the 355-bp region (displayed as a red rectangle) analyzed for methylation and overlapping the CG (cg14892768) identified by the array. Integration of placenta and sperm specific methylation values (Rakyan et al. 2008) into the Ensembl Genome Browser (www.ensembl.org). Green and blue represent intermediate and methylated regions, respectively. (B) Detailed sequence information of the region analyzed for methylation surrounding the AXL promoter region. (C) The 14 CGs analyzed are shown in bold in B and were assayed by standard bisulfite sequencing; the black arrow represents the same underlined CG in B and corresponds to the CG identified by the array. Filled black circles represent methylated CG and open circles represent unmethylated CG. (D) Direct bisulfite sequencing of a subset of CG sites marked by * in B in the same placental sample as C. (E) Box-and-whisker plot displaying the distribution of bisulfite pyrosequencing median percent CpG methylation for four consecutive CpG sites marked ^# in B within the new candidate AXL DMR for each tissue. The bisulfite pyrosequencing data are derived from 15 blood samples, 10 placenta samples, three AnCHM samples, and one MCT sample.