1. Expression of osteoactivin in rat and human liver and isolated rat liver cells 
Borislava Haralanova-Ilieva, Giuliano Ramadori, Thomas Armbrust 
Journal of Hepatology 
Volume 42, Issue 4, Pages (April 2005)
DOI: /j.jhep
Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

2. Fig. 1
Expression of osteoactivin (OA) in rat organs. Total RNA was extracted from rat organs and tissue and analysed by RT-PCR (A) or Northern blotting (B). (A) 1μg of RNA were reverse transcribed and subjected to PCR using OA-specific or GAPDH-specific primer pairs. PCR reactions were analysed by agarose gel electrophoresis revealing the expected 1.7kb PCR-product of OA (upper panel) or the 360bp PCR-product of GAPDH (lower panel), M-100bp DNA ladder. (B) 5μg of total RNA were separated on formaldehyde/agarose gel, transferred onto Hybond membrane and successively hybridised with radiolabelled, OA-specific probe (upper panel, detecting the single 2.3kb band of OA mRNA), or 28S rRNA-specific probe (lower panel).
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

3. Fig. 2
Normal (A–D) and acutely damaged liver (E–H) obtained 48h after CCl4 administration were cryocut (10μm slices). Nonradioactive ISH was performed using a digoxigenin-labelled, specific osteoactivin (OA) sense (A, C, E, G) or antisense (B, D, F, H) probe. Hybridised probes were detected by alkaline phosphatase-labelled sheep antibody directed against digoxigenin. Immunocomplexes were then detected with the NBT/BCIP yielding dark blue cytoplasmatic staining. Sections from normal (C, D) or acutely damaged (G, H) liver were stained thereafter with a monoclonal anti-CD68 antibody. Reaction was visualised using AEC revealing dark red cytoplasmatic reaction (Original magnification ×200).
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

4. Fig. 3
Expression of osteoactivin (OA) in isolated rat cells. (A) RT-PCR analysis of OA (upper panel) or GAPDH (lower panel) in isolated liver cells—hepatocytes (HEPs), small Kupffer cells (sKCs), large Kupffer cells (lKCs) at days 0 and 1 of culture, sinusoidal endothelial cells (SECs) and rat myofiroblasts (rMFs), M 100bp DNA ladder. (B) 2.3kb band of OA mRNA in HEPs at day 1 of culture, SECs at day 0, small KCs at day 0, large KCs at days 0, 1 and 5 of culture, myofibroblasts (rMF), hepatic stellate cells (HSCs) and rat peritoneal macrophages (PM). Lower panel: 28S rRNA. (C) The 422bp PCR product of OA in isolated rat osteoblasts and in cells from the mononuclear phagocyte lineage: rat osteoblasts (OBs), peripheral blood lymphocytes (PBLs), monocytes (Mo), Kupffer cells (KCs), peritoneal macrophages (PM), M 100bp DNA ladder. Lower panel: the 360bp control PCR product of GAPDH. (D) Densitometric analysis of the expression of OA in KCs with increasing time of culture corresponding to slots 3–6 of panel B.
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

5. Fig. 4
Northern blot analysis of osteoactivin (OA) expression in acute liver injury induced by CCl4. Total RNA was extracted from livers of control rats or 6, 12, 24, 48, 72 or 168h after one single oral dose of CCl4. RNA was electrophorised, transferred onto Hybond membrane and successively hybridised with radiolabelled, OA-specific (upper panel) or 28S rRNA-specific probe (lower panel). OA mRNA steady state level increased 12h after CCl4, peaked 48h after CCl4 and returned to baseline levels 168h after CCl4 administration.
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

6. Fig. 5
Expression of osteoactivin (OA) in isolated mononuclear phagocytes (MNP) of acutely injured rat liver. (A) Rats were given CCl4 and MNP isolated from control animals and from animals 1, 4, 12, 24, 48 or 72h after CCl4 treatment. The 2.3kb band indicates OA mRNA expression that increased 12h after CCl4, peaked at 48h and decreased thereafter. Control hybridisation with 28S rRNA (lower panel). (B) Densitometric analysis of OA expression in inflammatory MNPs.
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

7. Fig. 6
Modulation of osteoactivin (OA) expression in isolated normal rat Kupffer cells. Cells were cultured for one day and then treated with dexamethasone (Dex) (10 and 100nM/ml), LPS (10 and 100ng/ml), interferon-γ (γIFN) (100 or 1000U/ml) or transforming growth factor-β (TGF-β) (1 and 10ng/ml). RNA was extracted and analysed by Northern blotting as described. Upper panel: OA, lower panel: 28S rRNA. Densitometric analysis of bands is shown to the right.
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

8. Fig. 7
Northern blot analysis of osteoactivin (OA) expression in inflammatory MNP isolated 48h after CCl4 administration and treated with dexamethasone (Dex), lipopolysaccharide (LPS), interferon-γ (γIFN), or transforming growth factor-β (TGF-β) as indicated in Fig. 6 (amounts per ml; c, control). Lower panels: 28S rRNA.
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2005 European Association for the Study of the Liver Terms and Conditions