Figure 4

(A) The sequence of CpG islands in TFPI-2 promoter. The CpG dinucleoide detected by bisulfite modified sequence was indicated in gray shadow. (B) The TFPI-2 gene promoter is highly methylated in MDA-MB-435 cells, whereas it is largely unmethylated in MCF-7 cells. For each cell type, the methylation status of 8 individual clones as determined by bisulfite sequencing analyses is shown in rows 1 to 8. The filled or open circles represent the methylated or unmethylated CpG sites, respectively. (C, D) MDA-MB-435 cells were seeded into 10 cm tissue culture dishes at an initial density of 33% confluence, allowed to attach over night, and then replaced with medium containing 1.25, 2.5 and 5 μM of AZA for 72 hour, then expression of TFPI-2 was detected by western-blot (C) and real-time PCR (D).