Clinical, biochemical and histopathological patient characteristics.

*<p><b>…steatosis present, albeit <5% of parenchymal area</b>.</p><p>Samples from the Biobank cohort analyzed by qRT-PCR (p-value: a. steatohepatitis vs. steatosis, b. steatohepatitis vs. control, c. steatosis vs. control).</p

Validation of <i>AKR1B10</i> (A and C) and <i>KRT23</i> (B and D) mRNA expression by qRT-PCR in biopsy (top) and biobank (bottom) samples.

<p><i>HPRT1</i> was chosen to normalize the gene expression in the analyzed samples. Mean normalized expression levels are given in log2.</p

Overview of samples applied to screening and validation of deregulated genes in steatohepatitis and steatosis.

<p>Sample from the screen were also included in the validation.</p

Candidate genes for qRT-PCR validation.

<p>FCs (fold changes) of selected target genes were calculated from gene expression array data for the Biobank cohort. Gene expression from selected target genes was validated by qRT-PCR in the Biobank cohort.. FC was calculated between steatohepatitis (SH) and controls (ctrl) as well as between SH and steatosis.</p

Gene Ontology (GO) analysis.

<p>Over-represented GO terms for up regulated genes in common gene list in steatohepatitis samples (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046584#pone-0046584-g001" target="_blank">Fig. 1A</a>, cluster 2).</p

Fold changes of selected genes validated by qRT-PCR.

<p>The expression of selected target genes was determined in surgically collected samples (A) and in liver samples obtained by biopsy (B). The fold change was calculated by comparing the three groups of liver samples against each other.</p

Clinical, biochemical and histopathological patient characteristics.

<p>Samples from the Biopsy cohort used for the validation by qRT-PCR (p-value: a. steatohepatitis vs. steatosis, b. steatohepatitis vs. control, c. steatosis vs. control).</p

Immunohistochemical detection of the expression of AKR1B10 in chronic hepatitis C (A,B), fatty liver (C,D) and (cirrhotic) steatohepatitis (E,F).

<p>Only representative areas are shown. (A) Case of chronic hepatitis C with an inflamed portal tract with lymphocytic infiltrates and mild interphase hepatitis (central vein marked by asterisk, H&E stained section). (B) Consecutive section of the area shown in (A). Only a group of few centrilobular hepatocytes exhibit cytoplasmic and nuclear AKR1B10 immunostaining (central vein marked by asterisk). (C) Case of fatty liver with marked macro- and mediovacuolar steatosis predominantly of centrilobular and mid-zonal hepatocytes (central vein marked by asterisk; H&E stained section). (D) Consecutive section of the area shown in (C) of the hepatocytes with fatty change show staining of the rim of cytoplasm not occupied by fat with AKR1B10 antibodies (central vein marked by asterisk). (E) Case of steatohepatitis in a cirrhotic liver with parenchymal nodule abuting a fibrous septum with mild ductular reaction. Many hepatocytes show fatty change and some of them are characterized by an enlarged, lightly stained cytoplasm (ballooned hepatocytes) and irregular eosinophilic cytoplasmic inclusions (Mallory-Denk bodies, MDBs; inset with higher magnification showing ballooned hepatocytes containing MDBs; H&E stained section). (F) Consecutive section of the area shown in (E). Many of the normal-sized as well as the ballooned hepatocytes show moderate cytoplasmic immunostaining with AKR1B10 antibodies whereas the MDBs remain unstained (inset with higher magnification shows ballooned hepatocytes with MDB).</p

Candidate genes for qRT-PCR validation.

<p>Gene expression from selected target genes was validated in the Biopsy cohort. FC was calculated between steatohepatitis (SH) and controls (ctrl) as well as between SH and steatosis.</p

Immunohistochemical expression of AKR1B10 in liver parenchyma of patients with steatohepatitis, steatosis and chronic hepatitis C.

*<p>Weak and moderate AKR1B10 expression (% of parenchymal area): 0: no expression; 1: 1–10%; 2:>10–30%; 3:>30%.</p>**<p>Marked AKR1B10 expression (% of parenchymal area): 0: no expression; 1: 1–10%; 2:>10–30%; 3:>30%.</p