A. Holo-Tf induces hepcidin expression in WIF-B cells. WIF-B cells were left untreated or incubated with 25 μM human apo-Tf or holo-Tf for 24 hours, prior to RNA isolation. The hepcidin mRNA level was measured by qRT-PCR and normalized to GAPDH. B. Comparison of HFE, TfR1, TfR2 and HJV mRNA levels in WIF-B cells, HepG2 cells, and rat hepatocytes. Levels of TfR1, TfR2, HFE and HJV mRNA, iron related genes upstream of hepcidin, were measured by qRT-PCR. All samples were run in triplicate in three independent experiments. Data are shown as average ± S.D. P-values < 0.05 are indicated by *.