Fig. 1: Rnf43/Znrf3 (R&Z) deletion induces hepatocyte proliferation, hepatomegaly, tissue degeneration and steatohepatitis.

a Experimental design. Liver-specific Rnf43/Znrf3^del mice (R&Z^del) were generated by injecting AAV8-TGB-Cre virus to 8–12week-old Rnf43/Znrf3^flox mice (R&Z^flox) or by injecting tamoxifen (Tam) to 8–12-week-old AlbCre-ERT2-R&Z^flox mice. Control R&Z^flox mice received AAV8-null virus or vehicle. Livers were collected at 1 week (1W), 3 months (3 M) or 7 months (7M) later. Red dot, blood collection. b R&Z^del results in hepatomegaly. The graph represents the percentage of liver-to-body weight ratio. Results are presented as mean + /− SD from R&Z^flox 1W, n = 5; R&Z^del 1W; R&Z^flox 3M, n = 7; R&Z^del 3M, n = 6; R&Z^del 7M, n = 9; R&Z^flox 7M, n = 10. Unpaired two-tail t test. 1W, ***p = 0.0007; 3M, **p = 0.0017; 7M, ***p = 0.0001. c Oil red-O staining in R&Z^del 3M. Representative pictures of n = 3 independent experiments. Scale bar, 100 μm. d NAS scoring. Each column represents an independent mouse. Pink = NAS score 0–2 = Normal; clear violet = NAS score 3–4= borderline; dark violet = NAS score 5–8= steatohepatitis. e Representative histopathological analysis showing nodules (dashed circle) with cellular degeneration and ballooned hepatocytes (arrowheads) in R&Z^del livers (n = 7). Scale bar, 100 μm (left) and 50 μm (right). f R&Z^del livers present a significant increase in the number of proliferating (Ki67+, arrowheads) cells. Representative pictures. Scale bar, 50 μm. Graphs represent the percentage of Ki67+ hepatocytes per field-of-view (FOV). Violin plot represents median, IQR and full distribution of all FOVs (n  = 10 per mouse). Dots, mean of all FOV per mouse (n  = 3). Unpaired two-tail t test of means, **p = 0.0011. g Representative pictures of liver sections stained for p21 (arrowheads, p21+ cells). Scale bar, 50 μm. The graph represents the quantification of p21+ cells at 7M of age. Data are represented as violin plots showing median, IQR and full distribution of FOVs (n  = 10). Dots, mean of all FOV per mouse (n  = 3). Unpaired two-tail t test of means, **p = 0.0034. h Cleaved Caspase-3 and p-MLKL staining in control (top) and R&Z^del (bottom). Representative images from n = 3 independent 7M old mice. Scale bar, 100 μm. i Serum levels for ALT and ALP. Data represent mean + /− SD of n = 5 R&Z^flox and n = 6 R&Z^del samples. Unpaired two-tail t test. ALT, *p = 0.0376; ALP, *p = 0.0279. Source data are provided as a Source data file.