Glycol-split heparins reduce hepcidin and Id1 mRNA in a dose-dependent manner. (A) HepG2 cells were treated for 16 hours with nonanticoagulant heparins RO-82, RO-68, NAc-91, and NAcRO-00 at different concentrations (0.12-0.4-1.2-3.6-11-33 μg/mL) and in the presence of 10 ng/ml BMP6 . Hepcidin (A) and Id1 mRNA levels (B) were quantified with qRT-PCR in a relationship to Hprt1 mRNA. Data are means and SD of 3 different experiments and are expressed as fold increase. Anticoagulant heparin was used as control. (C) Western blotting analysis of phosphorylated SMAD1/5/8 and of Actin (as calibrator of the cell extracts) after the incubation with BMP6 and heparins. The image is representative of 3 independent experiments.