Glycol-split heparins reduce Oncostatin-M and IL-6-induced hepcidin mRNA and pSMAD but not pSTAT3. (Top) HepG2 cells were treated for 6 hours with 3.6 μg/mL glycol-split heparins RO-82 and RO-68 alone (solid histograms) or in combination with 2 ng/mL Oncostatin-M (empty histograms) and 50 ng/mL IL-6 (striped histograms). Hepcidin mRNA levels were quantified with qRT-PCR in relationship to Hprt1 mRNA. The values are presented as means and SD of 3 different experiments and are expressed as fold of increase of untreated cells. (Bottom) Western blotting analysis of pSMAD1/5/8, pSTAT3, and Actin. Actin was used as a calibrator of the cell extracts. The image is representative of 3 different experiments.