Heparanase Overexpression Reduces Hepcidin Expression, Affects Iron Homeostasis and Alters the Response to Inflammation
Fig 1
HepG2 cells transiently transfected with heparanase showed a reduction of hepcidin mRNA.
HepG2 cells were transfected with pcDNA3.1-HPA plasmid (HPA) or empty pcDNA3.1 as control (MOCK) and harvested 48 h after the transfection. (A) Relative level of HPA mRNA was measured by qRT-PCR (B) Western blot of SDS-PAGE with anti-HPA antibodies show the levels of its latent (65 kDa) and active (50 kDa) form. Densitometry quantification of the two protein forms was performed in relation to Actin. (C) The level of hepcidin mRNA and (D) Id1 mRNA was analyzed by qPCR and normalized for Hprt1. (E) The phosphorylated (pSMAD5) and total SMAD5 were analyzed by western blot and pSMAD5 densitometry was normalized to actin. In (A) the values are expressed as–dCt for HPA mRNA, in C and D as fold change over the control (MOCK) for hepcidin and Id1 mRNA., respectively
