Figure 2.

Hypermethylation of CpGs mediated decreased SNORD113-1 expression. A. Demethylation following treatment with 5-aza-2′-deoxycytidine induced SNORD113-1 upregulation. Left panel: a representative result; Right panel: the results from three experiments. B. Upper panel: a schematic of the Dlk1-Dio3 region in 14q32 [10]; Lower panel: the CpG island enriched sites in the upstream region of SNORD113-1 were analyzed using CpG island searcher, CoreBoost_HM and MethPrimer. C. Strong promoter activity of fragment 1 (F1) was demonstrated using luciferase assays. D. Bisulfite sequencing of the putative promoter region. Left panel, each line represents one PCR product, and five PCR products are shown for each sample. ●, methylated CpGs; ○, unmethylated CpGs. Right panel, the methylation status of each CpG in this region was quantified using the percentage of methylated CpGs among all PCR products. *p < 0.05, **p < 0.01.