Figure 5. Association between CpG methylation and the expression of HOXC10. (A) Total cell DNA was extracted from A549 and NCI-H23 cell cultures. Cytosine methylation in the HOXC10 CpG island was compared between two culture conditions using bisulfite treatment coupled with methylation-specific qPCR. A fold change of the methylated and unmethylated PCR products were obtained by setting the values from 2D culture to one. (B) Total cell RNA was extracted from A549 cells exposed to either a DNMT inhibitor, 5-Aza-CdR (1 μM) or DMSO for 72 hrs. The RNA levels of HOXC10 were measured using qRT-PCR. A fold change was obtained by normalizing to the housekeeping gene RPLP0 and setting the values from the DMSO group to one. When presented, means and standard deviations were obtained from at least 3 independent experiments. * and ** indicate a P value < 0.05 and 0.01, respectively.