Effect of PG-APS administration and phlebotomy on serum hepcidin levels and liver hepcidin mRNA expression. Serum hepcidin (A) was determined in control, IDA, ACD, and ACD/IDA rats by liquid chromatographic separation and tandem mass spectrometry detection. For determination of liver hepcidin mRNA (B), nitrogen snap-frozen tissue was subjected to RNA preparation, followed by reverse transcription and quantitative TaqMan PCR and normalized to the mRNA expression levels of the housekeeping gene β-glucuronidase (Gusb). Figure 1 contains details on graphs and statistics.