Hematological indices and hepcidin mRNA levels in PHZ-treated mice. (a) Hematological parameters: RBC (106/ml), HGB concentration (g/dl), HCT (%), and MCV (fl). Results are expressed as mean ± SD for n animals. (b) Hepcidin mRNA content in liver of control and PHZ-treated mice 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). Autoradiography of a typical experiment. (c) 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 < 0.0001. (d) Level of specific hepc1 transcripts in the liver as measured by RT-PCR (as described in Methods). Following PCR, the amplified products (171 bp for hepc1 and 250 bp for β-actin) were separated by electrophoresis on 1.5% agarose gel.