Figure 6.

Effect of 27nt RNA duplex on CpG island methylation in eNOS gene. (A) Methylation status in CpG islands in the genomic region immediate upstream of the eNOS intron 4. Sodium bisulfite genomic sequencing was used for the experiments in HAECs treated with 27 RNA duplexes or siLuc (negative control) for 24 h. The x-axis indicates the CpG island position in reference to the transcription starting site of the eNOS gene. Each column represents the percentage of the methylated CpG of a total of 10–15 individual clones. An independent Student's t test was performed to compare the differences between siLuc siRNA and 27nt RNA treated cells. (**p < 0.01). The quantitative data represent the results of three independent experiments. (B) Western blot of the eNOS, DcR1 and ICAM-1 proteins in HAECs treated with 27nt RNA duplex with or without the 5-aza–methyl transferase inhibitor. 5-aza seemed to recover the 27nt RNA duplex-induced eNOS depression in HAECs.