Fig. 2

Effects of 5-Aza-CdR treatment on MMP-9 CpG methylation and H3NT proteolysis. a Mock-depleted or MMP-9-depleted OCP-induced cells were cultured with or without 5-Aza-dC for 0, 1, 3 and 5 days. Chromatin was extracted, and the extent of H3NT proteolysis was analyzed by Western blotting with H3 C-terminal tail antibody. H2A was used as a loading control. b The sequence of the MMP-9 CpG island locus (NCBI accession: NC_000068, region: 164950229 to 164950544) containing the 16 CpGs that are written in bold and underlined. c Genomic DNA was obtained from OCP-induced cells that were grown in the presence or absence of 5-Aza-CdR for 5 days. The purified DNA molecules were subjected to sodium bisulfite conversion reaction and then analyzed by methylation-specific PCR of the MMP-9 CpG island. M, methylated; U, unmethylated. d Bisulfite sequencing analysis of the MMP-9 CpG island locus using genomic DNA prepared as in c. White circles, unmethylated CpGs; black circles, methylated CpGs