Targeted Disruption of the Hepatic Transferrin Receptor 2 Gene in Mice Leads to Iron Overload  Daniel F. Wallace, Lesa Summerville, V. Nathan Subramaniam  Gastroenterology  Volume 132, Issue 1, Pages 301-310 (January 2007) DOI: 10.1053/j.gastro.2006.11.028 Copyright © 2007 AGA Institute Terms and Conditions

Figure 1 Measurement of iron indices in LSKO and complete TfR2-KO mice. Mean values for transferrin saturation (A, B), hepatic iron concentration (C, D), and splenic iron concentration (E, F) are shown for 5-, 10-, and 26-week-old control (white bars) and KO (shaded bars) mice. Controls and LSKO mice: A, C, and E; controls and complete TfR2-KOs: B, D, and F. Error bars denote standard error of the mean; **, P < .01. Gastroenterology 2007 132, 301-310DOI: (10.1053/j.gastro.2006.11.028) Copyright © 2007 AGA Institute Terms and Conditions

Figure 2 Perls’ staining of liver sections. Representative liver sections from 10-week-old control (A) and LSKO (B) mice stained with Perls’ Prussian blue for iron, showing predominantly periportal accumulation of iron in hepatocytes of the LSKO mouse. Original magnification 20X. Gastroenterology 2007 132, 301-310DOI: (10.1053/j.gastro.2006.11.028) Copyright © 2007 AGA Institute Terms and Conditions

Figure 3 Immunohistochemical localization of TfR2 in the liver. Liver sections from 10-week-old control (A, B) and LSKO (C, D) mice were incubated with anti-TfR2 antibodies (A, C) or control rabbit IgG (B, D). Staining for TfR2 expression is observed on the basolateral surface of hepatocytes in livers from control mice. No staining is observed in LSKO mice or with rabbit IgG. Original magnification 40X. Gastroenterology 2007 132, 301-310DOI: (10.1053/j.gastro.2006.11.028) Copyright © 2007 AGA Institute Terms and Conditions

Figure 4 Quantification of hepatic mRNA expression in 10-week-old mice. The relative expression of mRNA transcripts in the livers of 10-week-old LSKO mice (A) and complete TfR2-KO mice (B) are shown relative to respective control levels. Error bars denote standard error of the mean. *, P < .05; **, P < .01. Gastroenterology 2007 132, 301-310DOI: (10.1053/j.gastro.2006.11.028) Copyright © 2007 AGA Institute Terms and Conditions

Figure 5 Quantification of hepatic hepcidin mRNA expression in 5-, 10-, and 26-week-old mice. The relative expression of hepcidin mRNA transcripts in the livers of 5-, 10-, and 26-week-old LSKO mice (A) and complete TfR2-KO mice (B) are shown relative to respective control levels. Error bars denote standard error of the mean. *P < .05. Gastroenterology 2007 132, 301-310DOI: (10.1053/j.gastro.2006.11.028) Copyright © 2007 AGA Institute Terms and Conditions

Figure 6 Quantification of hepatic protein levels. Total liver protein from 5-, 10-, and 26-week-old mice was Western blotted with antibodies against TfR2 (A), TfR1 (B), ferritin (C), and prohepcidin (D). Protein levels were quantified after normalizing to β-actin. Mean values are shown for control (white bars) and LSKO (shaded bars) mice. Error bars denote standard error of the mean. *, P < .05; **, P < .01. Gastroenterology 2007 132, 301-310DOI: (10.1053/j.gastro.2006.11.028) Copyright © 2007 AGA Institute Terms and Conditions