Figure 4.
HBP inhibits intracellular ferritin accumulation. (A-B) To assay intracellular iron sequestration in cultured cells, intracellular ferritin accumulation was measured in HepG2 cells (A) and FPN-EGFP–expressing COS-1 cells (B). HepG2 cells were maintained in minimum essential medium (Thermo Fisher Scientific) supplemented with 10% FBS and nonessential amino acids (Lonza). Cells were incubated with hepcidin (0.5μM). Cells were incubated with hepcidin (0.5μM) plus HBP (5.0μM). Cells were incubated with hepcidin (0.5μM) plus control peptide (5.0μM). Peptides and hepcidin were simultaneously added to the cell media. Bars represent means ± standard deviation (SD; n = 3). ∗P < .05. (C) Intracellular iron sequestration in an in vivo system. Hypoferremia was induced by hepcidin in mice. To suppress endogenous hepcidin, mice were fed a modified AIN-93G rodent diet with no added iron (Research Diet Inc) for 3 days. Mice received an intraperitoneal injection of hepcidin, hepcidin plus HBP, and hepcidin plus a control peptide. Peptides (900 nmol/kg body weight) and hepcidin (90 nmol/kg body weight) were separately administered to mice at the same time. Four hours later, the serum ion level was determined (n = 5-7). Comparisons between experimental groups were conducted with the use of multiple Student t tests. ∗P < .05. cont., control peptide.

HBP inhibits intracellular ferritin accumulation. (A-B) To assay intracellular iron sequestration in cultured cells, intracellular ferritin accumulation was measured in HepG2 cells (A) and FPN-EGFP–expressing COS-1 cells (B). HepG2 cells were maintained in minimum essential medium (Thermo Fisher Scientific) supplemented with 10% FBS and nonessential amino acids (Lonza). Cells were incubated with hepcidin (0.5μM). Cells were incubated with hepcidin (0.5μM) plus HBP (5.0μM). Cells were incubated with hepcidin (0.5μM) plus control peptide (5.0μM). Peptides and hepcidin were simultaneously added to the cell media. Bars represent means ± standard deviation (SD; n = 3). ∗P < .05. (C) Intracellular iron sequestration in an in vivo system. Hypoferremia was induced by hepcidin in mice. To suppress endogenous hepcidin, mice were fed a modified AIN-93G rodent diet with no added iron (Research Diet Inc) for 3 days. Mice received an intraperitoneal injection of hepcidin, hepcidin plus HBP, and hepcidin plus a control peptide. Peptides (900 nmol/kg body weight) and hepcidin (90 nmol/kg body weight) were separately administered to mice at the same time. Four hours later, the serum ion level was determined (n = 5-7). Comparisons between experimental groups were conducted with the use of multiple Student t tests. ∗P < .05. cont., control peptide.

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