Data files

Supplmentary Materials, Methods, Figures and Tables

List of used antibodies

Publications in which periportal fibrosis has been reported after repeated doses of CCl4.

Materials and Methods

PBC scoring of liver fibrosis in 12 to 84 weeks old Abcb4ko mice. Correlation analysis is based on Test z versus 12-weeks-old mice (PPTX 69 KB)

Abcb4ko mouse age, gender and number. Experiment 1 was used to investigate chronic liver disease development and progression. Experiment 2 was performed to test the anti-fibrotic efficiency of Galunisertib. (PPTX 36 KB)

Negative controls for IHC as used in this study. In the same staining batch, negative controls by omitting first antibodies were included. Scale bars are 100 µm. (PPTX 5991 KB)

Body and liver weight as well as liver enzymes upon Galunisertib treatment in 26-weeks-old Abcb4ko mice. a–c mouse and liver weight, and liver as percentage of total body weight. No significant difference is present due to Galunisertib administration. d–f) liver enzymes in Galunisertib, vehicle and untreated mice. e) H&E staining across different groups. Scale bars are 200 µm. Data are means ± SD of 5–7 mice per group. N.S.: P > 0.05; *: P < 0.05; **: P < 0.01 compared with untreated ...

Fig. S1 Liver cell proliferation, resident macrophage and leucocyte numbers upon Galunisertib treatment in 26-weeks-old Abcb4ko mice. a) Representative images of Ki-67 (proliferation marker), F4/80 (resident macrophages) and CD45 (leukocyte common marker) positive signals in liver tissues upon Galunisertib or vehicle administration. Scale bars are 100 µm. b and c) show numbers of proliferating hepatocytes and non-parenchymal cells, as indicated by Ki-67 positivity. d and e) quantification of F4/80 ...

Inducible Cre system and fate tracing cells using tamoxifen

Representative NMR spectra of different metabolites across the groups. and c) NMR-based metabolomics heatmaps of liver tissue revealed different patterns upon TAM administration. Red and green colors refer to upregulated and downregulated genes, respectively. The p value indicates the statistical difference between the presented groups (JPG 2849 KB)

Heatmap shows deregulated phase I and phase II metabolic targets as well as transporter () and inflammatory assigned (b) genes as quantified by the Fluidigm platform in TAM/Oil and Veh/Oil-exposed mice. Red and green colors refer to upregulated and downregulated genes, respectively. The p value indicates the statistical difference between TAM/Oil and Veh/Oil groups (JPG 3455 KB)

CYP2E1 activity in HepG2-E47 was measured. TAM inhibits (not significantly) the CYP2E1 activity compared to alcohol and DMSO-treated cells. C34 cell line was used as a negative control. Bars represent the means±SD of 3 biological replica. Student’s t test (paired t test) was used to check the statistical differences upon TAM incubation (JPG 1530 KB)

Protein level of CYP2E1 was analyzed by co-immunofluorescence staining immediately after attachment (3h) and 24 hours later. b and c) Protein and mRNA levels of CYP2E1 were analyzed upon TAM pretreatment. d and e) Protein and mRNA levels of catalase were assessed upon TAM pretreatment. Bars represent the means±SD of 3 mice (JPG 3000 KB)

Formalin-fixed livers were processed and stained with HE, CYP2E1, GS, F4/80 and CD45 to visualize and quantify hepatocellular necrosis, metabolizing enzymes, resident macrophages and recruited immune cells, respectively. Slides were scanned and the signal was randomly quantified in 15 images per liver. Yellow dashed lines indicate the necrotic areas. Yellow arrows refer to the positive signals. Scale bars are 200µm for HE and CYP2E1 and 100µm for GS, F4/80 and CD45 (JPG 6105 KB)