Generation of conditional KO mice for hepcidin. (A) Scheme of the mouse WT hamp1 genomic sequence (i), preceded by the usf2 gene loxP-flanked allele with FRT-flanked Hygro sequence (ii), floxed allele after flippase recombination (iii), and conditional KO allele after Cre-mediated excision (iv). Arrows: P1 and P2 primers used for deletion verification. (B) Recombination of the floxed hamp1 allele by genomic PCR in the pancreas, duodenum, spleen, liver, kidney, lung, testis, and heart of the Hepc^∆total, Hepc^∆liver and WT mice. (C) Hepcidin messenger RNA (mRNA) expression in the liver of Hepc^∆total and Hepc^∆liver mice and control littermates. n > 5. Values of WT mice were set up at 1. Samples were normalized to the threshold cycle value for cyclophilin. ***P < .001.