Hepatocyte Tfrc knockout mice (Tfrcfl/fl;Alb-Cre+) exhibit hypoferremia, reduced liver iron, and increased erythropoietic drive, without appropriate hepcidin suppression. Eight-week-old male (blue) and female (red) hepatocyte Tfrc knockout mice (Tfrcfl/fl;Alb-Cre+) and littermate controls (Tfrcfl/fl;Alb-Cre−) were analyzed for (A) serum transferrin saturation and (B) liver iron levels by colorimetric assays. (C) Serum EPO and ERFE protein levels were quantified by ELISA. (D) Livers were analyzed for Hamp relative to Rpl19 mRNA by qRT-PCR. (E) Livers were analyzed for pSMAD5 relative to total SMAD5 and actin protein by immunoblot and chemiluminescence quantitation. A representative immunoblot is shown. For panels D-E, the average of the control male mice was set to 1. (F) Hamp/Rpl19 mRNA was divided by liver iron content or serum iron content to normalize for the degree of iron overload and, (G) Hamp/Rpl19 mRNA was multiplied by serum EPO or ERFE levels to normalize for the degree of increased erythropoietic signals. For all graphs, individual data points are shown, and bars represent mean ± SEM. ∗P < .05, ∗∗P < .01, ∗∗∗ P < .001 relative to sex-matched Tfrcfl/fl;Alb-Cre− mice by the Student t test. ELISA, enzyme-linked immunosorbent assay; qRT, quantitative reverse transcription; SEM, standard error of the mean.
