Changes of iron metabolism gene expression in liver, spleen, and duodenum between the different anemia groups. Rats were inoculated on day 0 with an intraperitoneal injection of PG-APS to induce anemia of chronic disease (ACD) or left untreated (control). One group of PG-APS-treated and control rats was phlebotomized, starting 1 week before death, to create a combination of ACD and iron deficiency anemia (ACD/IDA) or IDA alone, respectively. Nitrogen snap-frozen tissue was subjected to RNA preparation, followed by reverse transcription and quantitative TaqMan PCR. In the liver (A), spleen (B), and duodenum (C), the TfR-1 (i), ferroportin (ii), and DMT-1 (iii) mRNA expression was determined by quantitative RT-PCR and normalized to the mRNA expression level of the housekeeping gene β-gluconidase (Gusb). Data are depicted as lower quartile, median, and upper quartile (boxes) and minimum/maximum ranges (whiskers). Calculations for statistical differences between the various groups were carried out by analysis of variance technique and Bonferroni correction for multiple tests.