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Figure 4. C. rectus infection induces hypermethylation of the placental-specific P0 promoter region of Igf2 in the placenta. DNA from unchallenged and from intra-uterine growth-restricted placentas were bisulfite-treated and sequenced (details in MATERIALS & METHODS). (a) The location of the CpG islands and the differentially methylated regions (DMR) of the Igf2 gene, as well as the location of the 4 promoters (P0, P1, P2, P3). (b) The 16 CpG dinucleotides (vertical gray lines) within the P0, which are also part of the DMR0. This DNA fragment extends from sites 7782 to 8512 (GenBank Accession #U71085). To facilitate the sequencing process, we divided this region into 3 areas (amplicons 1–3). (c) For each CpG locus, from 50 to 90 clones were sequenced per experimental group, giving a total of 390 clones. The 6 red vertical lines represent the CpG sites that were hypermethylated by more than 10% after C. rectus infection. *The CpG sites where hypermethylation was statistically significant (p < 0.05). (d) Depicts the percentage of methylation at each of the CpG sites from the unchallenged (red) and challenged with Intra-uterine Growth Restriction (blue) placentas. Mean % methylation and error bars are presented. Statistical analysis can be found in MATERIAlS & METHODS. *CpG sites where hypermethylation was statistically significant (p < 0.05). OR, odds ratio; parentheses include the confidence interval. Four CpG sites were significantly hypermethylated (7845, 7869, 8461, and 8472), and these sites appear to be clustered.
J DENT RES, Vol. 86, No. 2,
169-174 (2007)
DOI: 10.1177/154405910708600212
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