Mdr2^−/−IKK2^Hep-KO mice show impaired growth and signs of jaundice.

<p>(A) Immunoblot of whole liver lysates from 3 representative mice for each depicted genotype. Tubulin serves as loading control. (B) Weight curve of male mice for the denoted genotypes. * p<0,05 for Mdr2^+/− IKK2^Hep-KO vs. Mdr2^−/−, ** p<0,001 for Mdr2^−/− IKK2^Hep-KO vs. Mdr2^−/−. (C) Macroscopic appearance of littermate male mice at 12 weeks of age. Genotypes as specified in the picture. (D) Necropsy of littermate mice from (C). White arrowheads: enlarged gall bladder in double knockout mouse. Black arrowhead: jaundiced peritoneal serosa in double knockout mouse vs. normal appearance of serosa in Mdr2^−/− and Mdr2^+/−IKK2^Hep-KO mice. The ruler indicates centimeters in C and D.</p

Histological analysis of the liver pathology in Mdr2^−/−IKK2^Hep-KO mice.

<p>(A) Representative images of Hematoxylin & Eosin (HE) stained livers from young (12-week-old) littermate male mice and old (27–30 weeks) mice with the specified genotypes. Insert: High magnification of corresponding liver sections. (B) Immunofluorescence staining of cytokeratin 19 (CK19) in old male mice. DNA was stained with DAPI. The dashed line marks the distance from one portal field to the next, showing extreme abundance of CK19 positive cells in the Mdr2^−/− IKK2^Hep-KO as compared to Mdr2^−/− or Mdr2^+/−IKK2^Hep-KO mice. PV portal vein. CV central vein. BD bile duct. Scale bars, 100 µm.</p

IKK2-deficient hepatocytes are more sensitive to bile acid induced apoptosis compared to wild type cells.

<p>(A) Representative images of DAPI stained primary WT and IKK2-KO hepatocyte cultures. Cells were left untreated or were stimulated with taurolithocholic acid 3-sulfate disodium salt (TLCS, 100 µM) for the indicated time periods. Arrowheads show apoptotic hepatocytes as indicated by smaller highly fluorescent nuclei with condensed chromatin. Scale bars, 100 µm. (B) Graphs showing the average proportion of apoptotic hepatocytes from 3 independent experiments. Mean values ± SEM shown. * p<0,05; ** p<0,01; n.s.: not statistically significant. (C) Immunoblot detection of cleaved Caspase-3. Primary hepatocytes were stimulated with glycochenodeoxy-cholate (GCDC, 50 µM, top) or TLCS (100 µM, bottom) for the indicated time periods. Tubulin serves as loading control.</p

Mdr2^−/−IKK2^Hep-KO mice develop severe fibrosis.

<p>(A) Representative images of Masson trichrome stained livers from 12-week-old littermate male mice and from 27–30 week-old male mice. (B) Representative images of Sirius Red stained livers from 12-week-old littermate male mice and from old adult male mice (27–30 weeks). Insert: High magnification of corresponding liver sections. Arrowhead: Pericellular fibers. Double arrowhead: Pericellular space free of stained fibers. Genotypes as in (A). (C) Quantification of fibrosis in mice with the depicted genotypes measured as percentage of Sirius Red positive area as a fraction of the total area of at least 10 high power fields per mouse. Left: young adult mice (males, 8–19 weeks), right: old adult mice (males, 20–42 weeks). Error bars indicate SEM. Scale bars, 100 µm.</p

Mdr2^−/−IKK2^Hep-KO mice exhibit increased liver damage, hepatocyte death and cholestasis.

<p>(A) Alanine aminotransferase (ALT), (B) alkaline phosphatase (AP), and (C) bilirubin levels in serum of adult male mice (8–26 weeks) with the indicated genotypes. Error bars indicate SEM. Number of mice: Mdr2^−/−IKK2^Hep−KO: n = 10, Mdr2^−/−: n = 7, Mdr2^+/−IKK2^Hep-KO: n = 7 and Mdr2^+/+IKK2^FL: n = 2. (D–E) Representative pictures of TUNEL (D) and PCNA staining (E) on sections of paraffin-embedded livers from mice with the indicated genotypes. The green signal in E corresponds to background auto-fluorescence and was used to visualize the general morphology of the liver tissues. Scale bars: 20 µm.</p

Major HGF-mediated regenerative pathways are similarly affected in human and canine cirrhosis-1

<p><b>Copyright information:</b></p><p>Taken from "Major HGF-mediated regenerative pathways are similarly affected in human and canine cirrhosis"</p><p>http://www.comparative-hepatology.com/content/6/1/8</p><p>Comparative Hepatology 2007;6():8-8.</p><p>Published online 31 Jul 2007</p><p>PMCID:PMC1971050.</p><p></p>titis (CH), lobular dissecting hepatitis (LDH), cirrhotic samples (CIRR). Detection of the 86 kDa STAT3 protein (A), detection of the 60 kDa PKB protein (B), detection of the 44/42 kDa ERK 1/2 protein shown in (C), and detection of the p38-MAPK protein shown in (D). Beta-actin was used as a loading control

Quantitative evaluation of the percentage of tissue area exhibiting CD34 or sma immunopositivity and of nuclear area exhibiting PR immunopositivity (LI, labeling index) in the ULP area.

<p>Each dot shows the value associated with one case. The results of the Kruskal-Wallis test and the post-doc pair comparison test are shown below.</p

Additional file 4: Figure S3. of Prognostic relevance of molecular subtypes and master regulators in pancreatic ductal adenocarcinoma

Subtype functional characterization. (a) Venn diagram for differentially expressed genes for the predicted subtype comparisons (limma, adjust.method = BH, padj.thr = 0.05, lfc.thr = 1). Expression heatmaps of up- and down-regulated genes in Exocrine subtype (b), QM-PDA subtype (c), and Classical subtype (d). Gene overlap between PDAssign genes and only genes specifically up-regulated in our predicted subtypes is shown in (e). Genes from the overlap are listed on each heatmap (b, c, d). (EPS 12073 kb

Human Pancreatic Cancer Contains a Side Population Expressing Cancer Stem Cell-Associated and Prognostic Genes

<div><p>In many types of cancers, a side population (SP) has been identified based on high efflux capacity, thereby enriching for chemoresistant cells as well as for candidate cancer stem cells (CSC). Here, we explored whether human pancreatic ductal adenocarcinoma (PDAC) contains a SP, and whether its gene expression profile is associated with chemoresistance, CSC and prognosis. After dispersion into single cells and incubation with Hoechst dye, we analyzed human PDAC resections specimens using flow cytometry (FACS). We identified a SP and main population (MP) in all human PDAC resection specimens (n = 52) analyzed, but detected immune (CD45⁺) and endothelial (CD31⁺) cells in this fraction together with tumor cells. The SP and MP cells, or more purified fractions depleted from CD31⁺/CD45⁺ cells (pSP and pMP), were sorted by FACS and subjected to whole-genome expression analysis. This revealed upregulation of genes associated with therapy resistance and of markers identified before in putative pancreatic CSC. pSP gene signatures of 32 or 10 up- or downregulated genes were developed and tested for discriminatory competence between pSP and pMP in different sets of PDAC samples. The prognostic value of the pSP genes was validated in a large independent series of PDAC patients (n = 78) using nCounter analysis of expression (in tumor <i>versus</i> surrounding pancreatic tissue) and Cox regression for disease-free and overall survival. Of these genes, expression levels of <i>ABCB1</i> and <i>CXCR4</i> were correlated with worse patient survival. Thus, our study for the first time demonstrates that human PDAC contains a SP. This tumor subpopulation may represent a valuable therapeutic target given its chemoresistance- and CSC-associated gene expression characteristics with potential prognostic value.</p></div

Representative images of staining for alpha-sma (marker for ULP IC) showing the increased thickness of the alpha-sma+ ULP IC area in MS and BPS bladders.

<p>The ULP IC area lies between the 2 dotted red lines. The alpha-sma+ cell-types under the ULP IC area are MM-fibres and perivascular smooth muscle cells (which were excluded from analysis); IC in the DLP are negative for alpha-sma. In BPS bladder heavy inflammatory infiltrate is laying in between the IC, while in MS bladder this infiltrate is less pronounced. U is urothelium, ULP IC equals upper lamina propria interstitial cell. Scale bars indicate 600μm.</p