Figure 5

Effect of miR34a transfection with HA-based nanoparticles on mitochondrial DNA (mtDNA) Methylation levels. Mitochondria was isolated from A549 and A549 DDP cells post-transfection with miR34a HA-NPs (50 nM; 24 hours) DNA was isolated from this mitochondria and global (A) 5-methylcytosine (5-mC) (B) 5-hydroxymethylcytosine (5-hmC) content were measured with equal amounts of mtDNA using 5-mC and 5-hmC enzyme linked immunosorbent assay (ELISA) assays. Blank NPs–Blank HA-PEI/HA-PEG nanoparticles, miR34a - Naked miR34a without encapsulation, miR34a HA-NPs–microRNA34a encapsulated in HA-PEI/HA-PEG nanoparticles. (C) miR34 a HA-NPs induced changes in site-specific methylation content for individual CpG sites on the promoter regions of D-loop on mt-DNA as determined by bisulfite conversion followed by pyrosequencing. Pos1–3 indicates individual CpG sites. Results are expressed in units (mean +/− standard error of the mean [SEM]); n = 6; *p < 0.05 compared to the corresponding control.