C/EBPα protein levels and binding to HAMP promoter are modulated by EPO in a dose-dependent manner. (A) Western blot analysis of C/EBPα levels in HepG2 cells after treatment with physiological and supraphysiological doses of rEPO, anti-EPOR antibody, or IL-6. β-Actin was used as a loading control (bottom panel). Numbers on the left of each panel correspond to the sizes of the MW marker bands. Figure shows a representative result of 3 experiments. Vertical lines have been inserted to indicate repositioned gel lanes. (B) Scanning densitometry of C/EBPα Western blot. Data are normalized for β-actin levels and are expressed as arbitrary units (AU). (C) qRT-PCR–ChIP analysis of the effect of rEPO and anti-EPOR on the binding of C/EBPα to the HAMP promoter. Figure shows the averages and SD of 2 experiments, each with 3 replicates. * signals a significant (P < .05) difference between EPO-treated samples and the nontreated control (one-way ANOVA); ** signals a significant (P < .05) difference between anti-EPOR–treated samples and the 2.5 U EPO–treated cells (one-way ANOVA).