Effects of noggin, anti-BMP2/4 antibody, ALK1/Fc, and msHJV on hepcidin expression and response to holotransferrin treatment. Primary mouse hepatocytes from wild-type C57BL/6J mice were treated with a combination of (A) 1 μg/mL noggin plus 30 μM apoTf/holoTf or 100 EU/mL LPS (n = 5 mice); (B) 20 μg/mL anti-BMP2/4 antibody plus 30 μM apoTf/holoTf (n = 5 mice); (C) 500 ng/mL ALK1/Fc plus 30 μM apoTf/holoTf (n = 7 mice); (D) 30 μg/mL msHJV plus 30 μM apoTf/holoTf or 100 EU/mL LPS (n = 5 mice). (E) Comparison of the effects of msHJV, noggin, anti-BMP2/4 antibody, and ALK1/Fc on the induction of hepcidin by holotransferrin versus apotransferrin (left panel) and LPS versus solvent control (right panel). Statistics are as follows: *P ≤ .005; #P < .03, paired t test, comparing (A) noggin treatment with control, (B) anti-BMP2/4 antibody treatment with control, (C) ALK1/Fc treatment with control, (D) msHJV treatment with control; **P < .005, 1-way RM ANOVA, comparing apotransferrin, holotransferrin, and LPS treatments to respective solvent controls (E). Means and standard deviations are shown.