Fig. 6

Silencing of CCND2 AS1 due to its promoter CpG methylation in cervical cancer. aCCND2 AS1 expression was restored when the HeLa and SiHa cells were treated with 5΄-Aza from 5 μM to 50 μM. b Diagram of the CCND2 AS1 gene promoter with the transcription start site (TSS) indicated. The short black double-headed arrow represents the location of the fragment detected by the MSP and BGS assay. c Methylation status of the CCND2 AS1 promoter was detected by Methylation-specific PCR in HeLa and SiHa cell lines. M showed methylation-specific primer amplification, and U showed unmethylation-specific amplification. d Bisulfite genomic sequencing analysis was performed spanning the CCND2 AS1 core promoter and exon 1 in HeLa cell line. Twenty-two individual CpG sites within the CCND2 AS1 promoter were sequenced. Each row represents a single sequence; white circle indicates unmethylated CpG sites and black circle indicates methylated CpG sites