Generation of Bmp6 floxed (Bmp6fl/fl) and Bmp6−/−global knockout mice. (A) Schematic representation of Bmp6 gene structure and the floxed Bmp6 allele. Cre-mediated excision of exons 5 to 7 encoding the mature Bmp6 protein generates a functionally null allele. Genotyping primer positions are indicated. (B) PCR of tail genomic DNA from Bmp6 floxed and global knockout mouse colonies shows the presence of wild-type (WT) (+), floxed (fl), and cre-excised (–) alleles. (C) qRT-PCR of liver Bmp6 relative to Rpl19 mRNA in the Bmp6+/+, Bmp6+/−, and Bmp6−/− mice (n = 7 to 11 males and 5 to 7 females per group; supplemental Table 2). Results are reported as mean ± standard error of the mean (SEM) of –ΔCt = –[Ct target mRNA – Ct Rpl19], with a higher –ΔCt indicating higher mRNA expression. Fold change in mRNA expression between groups is calculated by 2–ΔΔCt, where ΔΔCt = ΔCt higher expressing group – ΔCt lower expressing group. *P < .05 relative to Bmp6+/+ mice by Student t test. nd, not detectable.