FIGURE 3.

K-Ras increases the MEN1 promoter DNA methylation via DNMTs. A, quantitative MS-PCR was used to analyze the methylation level of MEN1 promoter in K-Ras knockdown A549 cells (K-Ras shRNA), compared with Luc shRNA control. *, p < 0.05 versus control. B, bisulfite sequence analysis of each clone from K-Ras shRNA or Luc shRNA A549 cells (10 clones/sample). C, Western bolt detection of DNMTs and EZH2 in K-Ras diminished A549 cells. D, a schematic representation of the MEN1 gene loci and amplicons used for ChIP assay. ChIP assay was performed using the antibody against DNMT1 in vector and K-Ras-transfected A549 cells. PCR was carried out using primers for each amplicon.