These authors contributed equally to this work.
Liver Failure/Cirrhosis/Portal Hypertension
Disruption of the growth hormone—Signal transducer and activator of transcription 5—Insulinlike growth factor 1 axis severely aggravates liver fibrosis in a mouse model of cholestasis†
Article first published online: 30 NOV 2009
Copyright © 2009 American Association for the Study of Liver Diseases
Volume 51, Issue 4, pages 1319–1326, April 2010
How to Cite
Blaas, L., Kornfeld, J.-W., Schramek, D., Musteanu, M., Zollner, G., Gumhold, J., van Zijl, F., Schneller, D., Esterbauer, H., Egger, G., Mair, M., Kenner, L., Mikulits, W., Eferl, R., Moriggl, R., Penninger, J., Trauner, M. and Casanova, E. (2010), Disruption of the growth hormone—Signal transducer and activator of transcription 5—Insulinlike growth factor 1 axis severely aggravates liver fibrosis in a mouse model of cholestasis. Hepatology, 51: 1319–1326. doi: 10.1002/hep.23469
Potential conflict of interest: Nothing to report.
- Issue published online: 26 MAR 2010
- Article first published online: 30 NOV 2009
- Accepted manuscript online: 30 NOV 2009 12:00AM EST
- Manuscript Accepted: 29 OCT 2009
- Manuscript Received: 22 JUN 2009
- Ludwig Boltzmann Society
- Austrian Science Fund (FWF). Grant Numbers: SFB F28, P19188-B05
- GEN-AU grant “Austromouse” (Austrian Ministry of Science and Research)
Additional Supporting Information may be found in the online version of this article.
|HEP_23469_sm_SuppTab1.doc||46K||Supporting Table 1. Primers used for qPCR analysis|
|HEP_23469_sm_SuppTab2.doc||33K||Supporting Table 2. qRT-PCR primers and references for ChIP analysis|
|HEP_23469_sm_SuppTab3.xls||9635K||Supporting Table 3.|
|HEP_23469_sm_SuppFig1.tif||3930K||Supporting Figure 1: STAT5 and STAT3 activation status. (A) IHC staining for total STAT5B showed increased nuclear staining in hepatocytes (arrowheads) and cholangiocytes (arrows) of Mdr2-/- livers. Liver sections from control and STAT5Δhep mice did not show positive staining; immune cell infiltrates around fibrotic bile ducts of Mdr2-/-;STAT5Δhep liver stained positive for STAT5 while hepatocytes (arrowheads) and bile ducts (arrows) were not stained. (B) Western blot for total STAT5B, tyrosine-phosphorylated STAT5 (pY-STAT5), total STAT3 and tyrosine-phosphorylated STAT3 (pY-STAT3). STAT5B protein was undetectable in STAT5Δhep and Mdr2-/-;STAT5Δhep liver lysates. Increased tyrosine-phosphorylation of STAT5 was detected in Mdr2-/- livers. STAT3 protein levels were unchanged in all four genotypes while increased tyrosine-phosphorylated STAT3 was detected in Mdr2-/-;Stat5Δhep livers. HSC70: heat shock complex 70.|
|HEP_23469_sm_SuppFig2.tif||15920K||Supporting Figure 2: Effects of STAT5 deletion on macroscopic liver appearance, bile acid homeostasis and fibrogenesis. (A) Livers from Mdr2-/-;STAT5Δhep mice are pale compared to control and Mdr2-/- livers. STAT5Δhep livers are yellowish due to fat deposition. (B) Western blot analysis showed decreased Ntcp protein in STAT5Δhep livers, lack of Ntcp protein in Mdr2-/-;STAT5Δhep liver lysates and lack of Oatp1 protein in STAT5 deficient livers. (C) mRNA expression of Ntcp, Oatp1, Fxr and Shp1 was decreased in STAT5 deficient livers, Cyp7a1 mRNA was unchanged (n=6/genotype). (D) qRT-PCR revealed increased mRNA contents of Tgfβ, TgfβR1, Pdgfβ, PdgfβR1, Timp1-2, Mmp2, 3, 13-15 in Mdr2-/-;STAT5Δhep mice (n=6/genotype). (E) Western blot for TGFβ. Protein levels were similar in liver lysates from all four genotypes. HSC70: heat shock complex 70.|
|HEP_23469_sm_SuppFig3.tif||37K||Supporting Figure 3: Elevated hydroxyproline content in Mdr2-/-;STAT5Δhep liver tissue (n=6/genotype).|
|HEP_23469_sm_SuppFig4.tif||3781K||Supporting Figure 4: (A) Increased numbers of ki67 positive hepatocytes in Mdr2-/-;STAT5Δhep liver sections.|
|HEP_23469_sm_SuppFig5.tif||2561K||Supporting Figure 5: Reduced EGFR protein levels in STAT5 deficient livers. (A) Western blot analysis demonstrated lack of EGFR protein in STAT5Δhep and Mdr2-/-;STAT5Δhep liver lysates. (B) Tissue sections of STAT5 deficient livers did not stain positive for EGFR in IHC.|
|HEP_23469_sm_SuppFig6.tif||673K||Supporting Figure 6: Heat map listing changes in apoptotic gene expression as determined by Affymetrix gene expression analysis. mRNA levels of Caspase 12 (Casp12), Casp4, Casp1 and Casp6 as well as tumor necrosis factor related apoptosis-inducing ligand (Trail) and Fas ligand (Fasl) were increased in Mdr2-/-;Stat5Δhep livers. Colour code indicates upregulated (red) or downregulated (blue) gene expression versus controls.|
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