Fig. 2

Expression analysis and methylation analysis of PAX6 in cell lines. PAX6 and GAPDH expression were analyzed by the RT-PCR in the normal control, the normal liver cell line THLE-3, and the HCC cell lines treated with or without DNMT inhibitors (5-aza-2′-deoxycytidine; 5DAC) (a). Promoter methylation of PAX6 and COL2A was analyzed by MS-PCR with methylation-specific primers in the normal control, THLE-3, and HCC cell lines treated with or without 5DAC. CpG methylated human genomic DNA and DNA extracted from normal peripheral blood lymphocytes (PBL) which were modified by sodium bisulfite to generate a positive control, 1/5 diluted positive control, 1/20 diluted positive control, and a negative control, respectively (b). PAX6 methylation was also analyzed by bisulfite sequencing in Mahlavu cells treated with or without 5DAC. Each clone is represented by a row, and 30 CpG sites are represented as circles. Black and white circles represent methylated cytosine and unmethylated cytosine, respectively. Gray regions indicate the CpG sites that the MS-PCR/Q-MSP primer set covered (c)