1. Volume 125, Issue 2, Pages 460-469 (August 2003)
Heme oxygenase-1 is an antifibrogenic protein in human hepatic myofibroblasts 
Liying Li, Pascale Grenard, Jeanne Tran Van Nhieu, Boris Julien, Ariane Mallat, Aı̈da Habib, Sophie Lotersztajn 
Gastroenterology 
Volume 125, Issue 2, Pages (August 2003)
DOI: /S (03)

2. Figure 1
Expression of HO-1 protein during chronic liver diseases. Representative distribution of HO-1 immunostaining on liver tissue sections obtained from biopsy specimens of (A) normal liver and (B) active cirrhosis. Arrows indicate representative immunostaining of mesenchymal cells within and at the edge of the fibrotic septa and asterisks indicate representative immunostaining of Kupffer cells. The insets show respective negative control staining. Magnification 400×. (C–E) Colocalization of HO-1 and smooth muscle α actin by double immunostaining (C, smooth muscle α actin; D, HO-1 positivity; E, merge image; magnification 630×).
Gastroenterology  , DOI: ( /S (03) )

3. Figure 2
Induction of HO-1 in human hepatic myofibroblasts by hemin. (A) Northern blot analysis of HO-1 mRNA induction in human hepatic myofibroblasts treated with 10 μmol/L hemin for various periods of time. Expression of 28S ribosomal RNA was checked to correct for variations in mRNA loading and transfer (n = 2). (B) Western blot analysis of HO-1 protein treated with 10 μmol/L hemin for various periods of time. Expression of β-actin was checked to correct for variations in protein loading and transfer (n = 3).
Gastroenterology  , DOI: ( /S (03) )

4. Figure 3
Induction of HO-1 in human hepatic myofibroblasts by 15-d-PGJ2. (A) Northern blot analysis of HO-1 mRNA induction in human hepatic myofibroblasts treated with 1 μmol/L 15-d-PGJ2 for various periods of time. A typical autoradiogram is shown. After quantification of the autoradiographic signals, results were normalized relative to 28S ribosomal RNA expression to correct for variations in mRNA loading and transfer. Results are the mean ± SEM of 3 experiments (B) Western blot analysis of HO-1 protein treated with 1 μmol/L 15-d-PGJ2 for various periods of time. (C) Western blot analysis of HO-1 protein treated with varying concentrations of 15-d-PGJ2 for 9 hours. (B, C) Typical autoradiograms are shown. After quantification of the signals, results were normalized relative to β-actin expression (n = 6).
Gastroenterology  , DOI: ( /S (03) )

5. Figure 4
Cyclopentenone prostaglandins induce HO-1 protein expression, whereas PPARγ ligands have no effect. (A) Western blot analysis of HO-1 protein induction in human hepatic myofibroblasts treated with 1 μmol/L 15-d-PGJ2, 20 μmol/L Δ12-PGJ2, 30 μmol/L PGJ2, and 50 μmol/L PGD2, PGE2, PGA2, and 15-d-PGA2 for 9 hours. (B) Western blot analysis of HO-1 protein treated with either 1 or 4 μmol/L 15-d-PGJ2, or 10 or 20 μmol/L ciglitazone or troglitazone for 9 hours. Expression of β-actin was checked to correct for variations in protein loading and transfer.
Gastroenterology  , DOI: ( /S (03) )

6. Figure 5
HO-1 induction by 15-d-PGJ2 and hemin leads to inhibition of collagen type I synthesis in human hepatic myofibroblasts. (A) Time dependence of 15-d-PGJ2 inhibition of collagen(I) α1 mRNA expression. Human hepatic myofibroblasts were treated for varying periods of time with 4 μmol/L 15-d-PGJ2. (B) Human hepatic myofibroblasts were pretreated with or without 5 μmol/L SnPP for 2 hours, and further treated with 2 or 4 μmol/L 15-d-PGJ2 or vehicle for 24 hours. Northern blot analysis of collagen(I) α1 and α2 mRNA expression is shown (n = 5). (C) Human hepatic myofibroblasts were pretreated with or without 5 μmol/L SnPP for 2 hours, and further treated with 10 μmol/L hemin. Northern blot analysis of collagen(I) α1 and α2 mRNA expression (n = 5). After quantification of the autoradiographic signals, results were normalized relative to 28S ribosomal RNA expression to correct for variations in mRNA loading and transfer. P < by 2-way ANOVA for 15-d-PGJ2 and hemin treatment. ∗P < 0.01 for 15-d-PGJ2 or hemin vs. treatment with SNPP.
Gastroenterology  , DOI: ( /S (03) )

7. Figure 6
Antiproliferative effects of 15-d-PGJ2 and hemin are mediated by HO-1 in human hepatic myofibroblasts. Human hepatic myofibroblasts were pretreated for 2 hours with or without 5 μmol/L SnPP, and were stimulated further with 5% human serum, together with either (A) 3 or 5 μmol/L 15-d-PGJ2 or (B) 5 or 10 μmol/L hemin. Inset: dose-dependent effects of SnPP. Cells were pretreated with varying concentrations of SnPP and further stimulated with 5% human serum, together with 5 μmol/L 15-d-PGJ2. 5-Bromo-2-deoxy-uridine incorporation was measured as described in the Materials and Methods section. Results represent the mean ± SEM of 3–9 experiments, and are expressed as percent of control. P < by 2-way ANOVA for 15-d-PGJ2 and hemin treatment. ∗P < 0.01 for 15-d-PGJ2 or hemin vs. treatment with SNPP.
Gastroenterology  , DOI: ( /S (03) )

8. Figure 7
Bilirubin, and not CO or iron, mediates the antifibrogenic properties of HO-1. (A) DNA synthesis in 5% human serum-stimulated cells. Human hepatic myofibroblasts were treated either with varying concentrations of bilirubin dichloromethane, or preincubated for 1 hour with 100 μmol/L desferrioxamine (DF) before the addition of 3 μmol/L 15-d-PGJ2. Results represent the mean ± SEM of sextuplicate determination, and are expressed as percent of control. P < by 2-way ANOVA. (B) Northern blot analysis of collagen(I) α1 and α2 mRNA expression in cells treated for 5 hours with varying concentrations of bilirubin (n = 4). After quantification of the autoradiographic signals, results were normalized relative to 28S ribosomal RNA expression to correct for variations in mRNA loading and transfer. P < by 2-way ANOVA.
Gastroenterology  , DOI: ( /S (03) )