1. Lack of gp130 expression in hepatocytes promotes liver injury1
Konrad L StreetZ, Torsten Wüstefeld, Christian Klein, Karl-Josef Kallen, Francois Tronche, Ullrich A.K Betz, Günther Schütz, Michael P Manns, Werner Müller, Christian Trautwein 
Gastroenterology 
Volume 125, Issue 2, Pages (August 2003)
DOI: /S (03)

2. Figure 1
Normal liver development and time course of hepatocyte-specific gp130 deletion. (A) Crossbreeding of gp130loxP and AlfpCre mice generated hepatocyte-specific gp130-deficient mice (AlfpCre/gp130flox). In gp130loxP mice, 2 loxP sites flank exon 167 coding for the transmembrane domain. In AlfpCre mice, expression of the Cre recombinase is controlled by an albumin promoter/enhancer and AFP enhancer construct. (B) PCR analysis of embryonic tissue at time points postgestation. The 3.2-kb fragment represents the undeleted and the 1.0-kb fragment the deleted gp130loxP locus at exon 16. Liver DNA was prepared at days 9 to 14 as indicated. Extrahepatic embryonic tissue at day 14 (14B) was prepared as control. (C) Southern blot analysis of liver DNA derived from 6-week-old mice. The 5.5-kb fragment represents the wt gp130 locus, the 3.3-kb fragment the undeleted, and the 0.4-kb fragment the deleted gp130loxP locus.
Gastroenterology  , DOI: ( /S (03) )

3. Figure 2
Impaired STAT3 activation, induction of acute phase genes, and Akt-phosphorylation in AlfpCre/gp130flox mice. (A) Ten micrograms of IL-6 was injected per mouse, and liver nuclear extracts were prepared at time points indicated. Two micrograms of liver nuclear extracts and a 32P-labeled STAT3 consensus oligonucleotide were incubated and separated. The arrowhead depicts the position of bound STAT3. AlfpCre (+, positive) or (−, negative) indicates the presence of Cre in gp130flox mice. (B and C) Liver RNA was prepared before and at time points after IL-6 stimulation. Twenty micrograms of RNA/lane were analyzed by Northern blot analysis, hybridized with 32P-labelled probes for haptoglobin, serum amyloid A-2 (SAA), and SOCS3 (C). AlfpCre (+) or (−) indicates the presence of Cre in gp130flox mice. (D) Western blot analysis of phospho-Akt (60 kilodaltons). AlfpCre/gp130flox and control mice were stimulated with 10 μg IL-6 per mouse and analyzed before and after injection at the time points indicated using an anti-phospho Akt antibody. Equal protein loading was confirmed by Coomassie staining.
Gastroenterology  , DOI: ( /S (03) )

4. Figure 3
Lack of acute phase gene expression in AlfpCre/gp130flox mice after Oncostatin M stimulation. (A) Four micrograms of OSM were injected per mouse, and liver nuclear extracts were prepared at time points indicated. Two micrograms of liver nuclear extracts and a 32P-labelled STAT3 consensus oligonucleotide were incubated and separated. The arrowhead depicts the position of bound STAT3. AlfpCre (+; positive) or (−; negative) indicates the presence of Cre in gp130flox mice. (B) Liver RNA was prepared before and at time points after OSM stimulation. Twenty micrograms of RNA/lane were analyzed by Northern blot analysis, hybridized with 32P-labelled probes for serum amyloid A-2 (SAA). AlfpCre (+) or (−) indicates the presence of Cre in gp130flox mice. (C) Mice were stimulated with 4 μg OSM/mouse; SAA serum levels were determined before and 6 hours after OSM injection.
Gastroenterology  , DOI: ( /S (03) )

5. Figure 4
STAT3 and SOCS3 activation is impaired in AlfpCre/gp130flox mice after LPS stimulation. (A) AlfpCre (+) and (−)/gp130flox mice were stimulated with 100 μg LPS/mouse. IL-6 serum levels were determined at time points indicated. (B) RNA was prepared before and at the indicated time points after LPS stimulation. A duplex RT-PCR was performed with primers detecting IL-6 and GAPDH-specific signals. The position of the IL-6 and GAPDH-specific transcripts is indicated. (C) One hundred micorgrams of LPS were injected per mouse, and liver nuclear extracts were prepared at the time points indicated. Two micrograms of liver nuclear extracts and a 32P-labelled STAT3 consensus oligonucleotide were incubated and separated. The arrowhead depicts the position of STAT3. AlfpCre (+) or (−) indicates the presence of Cre in gp130flox mice. Super shift experiments were performed with an anti-STAT3 antibody. (D) Liver RNA was prepared before and at time points after LPS stimulation. Twenty micrograms of RNA/lane were analyzed by Northern Blot analysis and hybridized with 32P-labelled probes for SOCS3. AlfpCre (+) or (−) indicates the presence of Cre in gp130flox mice.
Gastroenterology  , DOI: ( /S (03) )

6. Figure 5
Gp130-dependent pathways control acute phase gene expression after LPS stimulation. (A) RNA was prepared from liver at time points after IL-6 stimulation. Twenty micrograms of RNA/lane were used for Northern blot analysis and hybridized with 32P-labeled probes for haptoglobin serum amyloid A-2 (SAA) and albumin. AlfpCre (+) or (−) indicates the presence of Cre in gp130flox mice. (B) Mice were stimulated with 100 μg LPS/mouse; SAA serum levels were determined at time points indicated.
Gastroenterology  , DOI: ( /S (03) )

7. Figure 6
Hepatocyte-specific gp130-deficient mice are more susceptible to LPS-induced liver failure. (A and B) Aminotransferases (ALT; A and AST; B) were determined before and at time points after LPS injection as indicated. AlfpCre (+) or (−) indicates the Cre status of gp130flox mice. (C) TUNEL assays of cryosections before and 24 hours after LPS injection in AlfpCre (+) and (−) gp130flox animals. (D) Quantification of TUNEL-positive liver cells after LPS challenge.
Gastroenterology  , DOI: ( /S (03) )

8. Figure 7
Reduced NF-κB activation despite higher TNF-α levels in AlfpCre/gp130flox mice after LPS stimulation. (A) AlfpCre (+) and (−) /gp130flox mice were stimulated with 100 μg LPS/mouse, and TNF-α serum levels were determined at time points indicated. (B) RNA was prepared before and at the indicated time points after LPS stimulation. A duplex RT-PCR was performed with primers detecting TNF and GAPDH-specific signals. The position of the TNF-α and GAPDH-specific transcripts are indicated. (C) One hundred micrograms of LPS were injected per mouse, and liver nuclear extracts were prepared at the time points indicated. Two micrograms of liver nuclear extracts and a 32P-labeled NF-κB consensus oligonucleotide were incubated and separated. The arrowhead depicts the position of NF-κB. AlfpCre (+) or (−) indicates the presence of Cre in gp130flox mice. Super shift experiments were performed with anti-p50 and anti-p65 antibodies as depicted. (D) Immunhistochemistry for NF-κB was performed in AlfpCre (+) or (−) gp130flox mice before and 1 hour after LPS stimulation as indicated.
Gastroenterology  , DOI: ( /S (03) )