Fig 3. CpG islands identified in ABCA1 promoter sequence are well conserved in evolution and ABCA1 hypermethylation occurs in plaque-laden aorta.

A, Observed/Predicted ratio of presence of CpG islands in ABCA1 promoter sequence (highlighted in yellow). Genomic position of promoter and ABCA1 CGIs obtained from different online websites (http://www.ebi.ac.uk/Tools/emboss/cpgplot/ and http://cpgislands.usc.edu/). B, Sequence alignment showing well conserved CpG islands and the binding sites of transcription factors. The conservation of ABCA1 CGI in some mammals obtained from the BLAST of NCBI (http://blast.ncbi.nlm.nih.gov/Blast.cgi?CMD=Web&PAGE_TYPE=BlastHome), and the transcription factor binding site is predicted by JASPAR (http://jaspar.genereg.net/). C, Hypermethylation of ABCA1 promoter in THP-1 and RAW264.7 macrophage-derived foam cells after EZH2 infection. PCR primers specific to unmethylated and methylated bisulfite-modified DNA were used to amplify the promoter of ABCA1 gene after EZH2 treatment. D, ABCA1 promoter methylation of the apoE^−/− mouse aorta, heart, liver and kidney was analyzed by MSP. UM: unmethylated; M: methylated.