JCI - Regulation of iron homeostasis by the hypoxia-inducible transcription factors (HIFs)

Regulation of iron homeostasis by the hypoxia-inducible transcription factors (HIFs)

Carole Peyssonnaux, Annelies S. Zinkernagel, Reto A. Schuepbach, Erinn Rankin, Sophie Vaulont, Volker H. Haase, Victor Nizet, Randall S. Johnson

Published in Volume 117, Issue 7

J Clin Invest. 2007; 117(7):1926–1932 doi:10.1172/JCI31370

Abstract | Full text | PDF

Options: View larger image (or click on image)

Figure 3

Binding of HIF-1 to the promoter of hepcidin and downregulation of hepcidin in Albumin-Cre/VHL^flox/flox.

(A) Sequence of murine (C57BL/6) hepcidin promoter; HREs are in bold; arrows indicate primers selected for ChIP. ChIP assay in vivo on liver extracts of WT and Albumin-Cre/VHL^flox/flox mice. (B) DFO (150 μM) induces binding of HIF-1 as shown by ChIP assay. (C) Luciferase-reporter constructs under the control of the regulatory region of the human hepcidin gene. HEK293 cells transiently transfected with pGL3 basic or pGL3-Hepc/HRE vector. (D) The “native” (CCACGTG) and mutated (CAA-TG) HREs (indicated by an X) are shown. HEK293 cells were transiently transfected with pGL3 basic, pGL3-Hepc/HRE, or pGL3-Hepc/mutHRE. (E) Hepcidin mRNA expression in livers of WT and Albumin-Cre/VHL^flox/flox by real-time RT-PCR (n = 8). HIF-1 and hepcidin expression in liver extracts of WT and Albumin-Cre/VHL^flox/flox mice. (F) Hepcidin mRNA expression in livers of WT, Albumin-Cre/VHL^flox/flox, and Albumin-Cre/VHL^flox/flox/ARNT^flox/flox (VHL^–/–ANRT^–/–) mice (n = 4). (G) IL-6 and IL-1β mRNA levels in livers of WT and Albumin-Cre/VHL^flox/flox mice.