The regulation of hepcidin expression does not involve DNA methylation.

<p>(A) Huh7.5-JFH1 cells were treated with or without 5-aza-2′-deoxycytidine (Aza), and hepcidin expression was analyzed by qRT-PCR. <i>GAPDH</i> was used as the internal control in the PCR reaction. The data are presented as mean ± SD from three independent experiments; n.s., not significant. (B) and (C) Genomic DNA extracted from Huh7.5, Huh7.5-JFH1 cells (B) or two paired liver tissues (C) was modified with sodium bisulfate, PCR amplified, and subsequently cloned and sequenced. The methylation status at each CpG site in CpG island of hepcidin promoter is shown. Methylated sites are indicated by filled dark circles and unmethylated sites by empty white ones. N: nontumor tissues; T: tumor tissues. (D) Huh7.5-JFH1 cells were treated with or without Trichostatin A (TSA), and hepcidin expression was analyzed by qRT-PCR. The data are presented as mean ± SD from three independent experiments.