Hepcidin mRNA level in transgenic mice constitutively expressinghepc2as determined by Northern blot and RT-PCR analyses. Twenty micrograms of total liver RNA purified from 66-day-old transgenic mice expressing hepc1¹¹  and from 21-day-old transgenic mice expressing hepc2 (A) or from 66-day-old nontransgenic (NT) mice compared with age-matched hepc2-transgenic (T) mice (B) were electrophoresed, blotted, and hybridized with the (³²P)-labeled hepcidin probe. This probe reveals both the transgenic hepcidin mRNAs (transgenic hepc1 mRNA and transgenic hepc2 mRNA, top band) and the endogenous hepcidin mRNAs. Note that in the 66-day-old hepc1-transgenic mice the endogenous genes (bottom band) are repressed or reduced (line 27) most likely due to, as reported,¹⁵  the hepc1-transgene-induced anemia. (C) Individual expression of hepc1 and hepc2 mRNAs was measured by RT-PCR, as described in “Materials and methods,” in 66-day-old nontransgenic (NT) mice compared with age-matched hepc2-transgenic (T) mice. Following PCR, the amplified products (171 bp for hepc1 or hepc2 and 250 bp for β-actin) were separated by electrophoresis on 1.5% agarose gel.