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Fig. S1. In vitro activity of recombinant HGF. Two canine cell lines were treated with recombinant canine HGF (Intervet, Boxmeer, the Netherlands, cHGF), recombinant human HGF (R&D Systems Europe, Oxon, UK, hHGF), and recombinant feline HGF (Zenoaq, Fukushima, Japan, fHGF). (A) Line graph representing the mean percentage growth of bile-duct epithelial cells (BDE) after 48 hours recombinant HGF treatment, as measured by the MTT assay. Cellular proliferation was induced at a concentration of 11 ng/mL HGF and a maximal proliferative activity was seen around 100 ng/mL HGF. No differences were seen in proliferative activity between recombinant fHGF, cHGF, and hHGF. (B) Effect of recombinant HGF treatment on the c-MET protein in BDE or Madin-Darby Canine Kidney cells (MDCK) after 5 minutes of treatment with HGF. Phosphorylation of c-MET was observed in both cell lines after 5 minutes of incubation with HGF. No differences were seen in c-MET phosphorylation after recombinant fHGF, cHGF, or hHGF treatment.

Fig. S2. Examples of Ki-67 positive hepatocytes. Immunohistochemical staining of Ki-67 (proliferation index) positive nuclei in hepatocytes of untreated (week 0) and after one week of recombinant HGF treatment are shown in (B). Original magnification × 200.

Fig. S3. ELISA measuring dog anti-fHGF IgG formation during and after HGF treatment compared to control. As no standardized concentration of dog anti-fHGF IgG was available, antibody formation was quantified as sample absorbance with subtraction of background signal. As a negative control, normal non-HGF treated dogs were used (NC). Sample absorbance retained basal levels in week 0 and 1, comparable to the negative control. However, in all dogs a clear increase in signal can be detected in week 2, 3, and 7 indicating anti-fHGF IgG formation during HGF treatment and persistence of circulating antibodies four weeks after treatment.

Table S1. Shunt index (%) calculated by perfusion scintigraphy before rHGF administration (week 0), during rHGF treatment (week 1 and 3) and four weeks after rHGF treatment (week 7).

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Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.