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The rest 25 methylated genes showed similar expression level between endosperm and embryo, among them, ten of the 25 genes were more methylated in embryo, the rest 15 genes showed similar methylation level between embryo and endosperm.
15 of the methylated genes showed much higher expression level in embryo than in endosperm (embryo/endosperm average RPKM >1.5), among them, one of the 15 genes was more methylated in endosperm, 6 were more methylated in embryo, and the rest 8 genes showed similar methylation level between embryo and endosperm.
Only one gene, DUSP4, displayed a significant difference in methylation level between supratentorial ependymomas and neural stem cells, being more methylated in the stem cells.
Figure 5 A shows a slightly bimodal distribution: The majority (72.7%) of CpG sites in all promoter regions across 28 tissues were unmethylated (methylation level ≤ 0.1), whereas 18.5% were semimethylated (methylation level between 0.1 and 0.7) and 8.8% were considered methylated (methylation level ≥ 0.7), according to the criteria established in Bell et al. (2011).
However, the molecular mechanisms behind the differences in DNA methylation level between the two groups of men were not immediately evident from the present results.
Of a total of 108 males, 30 samples were subjected to 450 K BeadChip arrays to evaluate the variation in DNA methylation level between cases and controls.
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These CpG sites were observed in higher frequencies in genes with methylation levels between 20 and 60% (Supplementary Fig. S3b).
Hemi-methylated CpG sites were also enriched in CpGs sites with methylation levels between 20 and 85% (Supplementary Fig. S3a).
Differences in methylation levels between embryo types were not significant for SOX2, LIN28 and GNAS.
To compare methylation levels between transfected samples, we tiled the genome with 200 bp tiles and performed paired t-tests for CpGs with ≥10 reads within the tile calculating p-values adjusted using the Benjamini Hochberg model.
In summary, the two types of demethylation should preferentially up-regulate genes that have 42 ~ 83 CpG sites near their TSS (−1.5 ~ + 2.0 kb) and methylation levels between 40 and 60%.
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