Volume 136, Issue 7, Pages 2316-2324.e3 (June 2009) Liver Zonation Occurs Through a β-Catenin–Dependent, c-Myc–Independent Mechanism  Zoé D. Burke, Karen R. Reed, Toby J. Phesse, Owen J. Sansom, Alan R. Clarke, David Tosh  Gastroenterology  Volume 136, Issue 7, Pages 2316-2324.e3 (June 2009) DOI: 10.1053/j.gastro.2009.02.063 Copyright © 2009 AGA Institute Terms and Conditions

Figure 1 (A) X-gal–stained liver from Ahcre mouse. (B–F) Hematoxylin and eosin staining of wild-type (B), Apc homozygous (C), β-catenin homozygous (D), Apc/β-catenin double homozygous (E), and c-Myc homozygous (F) liver sections. Gastroenterology 2009 136, 2316-2324.e3DOI: (10.1053/j.gastro.2009.02.063) Copyright © 2009 AGA Institute Terms and Conditions

Figure 2 Complementary changes in the zonation of GS and CPS 1 in the liver after β-naphthoflavone induction of AhCre+Apcfl/fl mice for 4 days. (A) RT-PCR analysis of cDNA from wild-type and AhCre+Apc+/fl and Apcfl/fl mice the perivenous markers GS, Glt1, RNAse4 and the periportal markers CPS, Arg1, Glutaminase 2, PEPCK, and GAPDH. (B) Western blotting analysis for total β-catenin protein, GS, CPS, Arg1, and GAPDH. (C) Immunostaining for β-catenin, GS, CPS, CYP2E1, and Lect2 on liver sections from mice with the indicated genotype. Gastroenterology 2009 136, 2316-2324.e3DOI: (10.1053/j.gastro.2009.02.063) Copyright © 2009 AGA Institute Terms and Conditions

Figure 3 Complementary changes in the zonation of GS and CPS I in the liver after β-naphthoflavone induction of AhCre+Apcfl/fl β-cateninfl/fl and β-cateninfl/fl mice for 21 days. (A) RT-PCR analysis. (B) Western blotting. (C) Immunostaining for perivenous and periportal markers as for Figure 2. Gastroenterology 2009 136, 2316-2324.e3DOI: (10.1053/j.gastro.2009.02.063) Copyright © 2009 AGA Institute Terms and Conditions

Figure 4 Disruption of zonation in Apc/c-Myc double homozygous liver occurs independent of c-Myc. Comparison of AhCre+Apcfl/+c-Myc+/+, Apcfl/+c-Mycfl/fl, and Apcfl/flc-Mycfl/fl liver by immunostaining after β-naphthoflavone induction for 21 days. Gastroenterology 2009 136, 2316-2324.e3DOI: (10.1053/j.gastro.2009.02.063) Copyright © 2009 AGA Institute Terms and Conditions

Supplementary Figure 1 Fold change in periportal and perivenous gene expression. Bar charts show the fold change in expression of CPS1, arginase1, GS, Glt1, RHBG, RNAse4, and β-catenin in wild-type AhCre+Apcfl/fl, AhCre+β-cateninfl/fl, AhCre+c-Mycfl/fl, AhCre+Apcfl/flc-Mycfl/fl, and AhCre+Apcfl/flβ-Cateninfl/fl animals. Statistically significant changes and P values are indicated (*). Gastroenterology 2009 136, 2316-2324.e3DOI: (10.1053/j.gastro.2009.02.063) Copyright © 2009 AGA Institute Terms and Conditions

Supplementary Figure 2 Fold change in periportal and perivenous protein levels. Bar charts show the fold change in the levels of CPS1, arginase1, GS, and β-catenin in wild-type AhCre+Apcfl/fl, AhCre+β-cateninfl/fl, AhCre+c-Mycfl/fl, AhCre+Apcfl/flc-Mycfl/fl, and AhCre+Apcfl/flβ-cateninfl/fl animals. Statistically significant changes and P values are indicated (*). Gastroenterology 2009 136, 2316-2324.e3DOI: (10.1053/j.gastro.2009.02.063) Copyright © 2009 AGA Institute Terms and Conditions

Supplementary Figure 3 Fold change in β-catenin target gene expression. Bar charts show the fold change in expression of Rrm2, Pfkfβ3, and Ihh in wild-type AhCre+Apcfl/fl, and AhCre+β-cateninfl/fl animals. Statistically significant changes and P values are indicated (*). Gastroenterology 2009 136, 2316-2324.e3DOI: (10.1053/j.gastro.2009.02.063) Copyright © 2009 AGA Institute Terms and Conditions