Effect of PHZ treatment in control mice and experimentally iron-overloaded animals. (a) Hepcidin mRNA content in liver of control mice, mice treated with PHZ, mice experimentally iron overloaded, and mice iron overloaded and treated with PHZ (see Methods) as determined by Northern blot analysis. Twenty micrograms of total liver RNAs were electrophoresed, blotted, and hybridized with both hepcidin and 18S-labeled probes (as described in Methods). Id, iron dextran. (b) Relative changes in hepcidin. mRNA level (18S normalized, arbitrary units) as calculated using a STORM850 PhosphoImager and ImageQuant 5.0. Results are expressed as mean ± SD for n animals, and statistical analysis was performed using the Student t test (unpaired, two-tailed). *P < 0.001 as compared with iron-overloaded mice.