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. 2020 Mar 30;27(5):1457–1474. doi: 10.1038/s41418-020-0532-1

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© The Author(s), under exclusive licence to ADMC Associazione Differenziamento e Morte Cellulare 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Micrographs of H&E stained liver sections from Otulin^∆hep and control mice aged 8–10 weeks. The dotted line indicates two subcapsular tumours. Micrographs are representative of eight mice of each genotype. Tu, tumour. b Quantification of the number of macroscopically discernible lesions (tumours, nodules, and discolourations) in Otulin^∆hep and control mice aged 8–10 weeks. Data are representative of four mice of each genotype. c Analysis of AFP levels in serum from terminal bleeds of Otulin^∆hep (n = 6) and control (n = 6) mice aged 8–10 weeks. d Relative mRNA expression of the indicated cancer markers in livers from Otulin^∆hep (n = 8) and control (n = 8) aged 8–10 weeks measured by quantitative RT-PCR. e Representative macroscopic appearance of Otulin^∆hep and control livers at the age of 50–54 weeks, grouped by severity. Arrowheads indicate highly vascularised tumours. Scale bars indicate 1 cm. f, g Quantification of the number of mice with the presence of a tumour ≥2 mm in diameter (f) or the number of mice in each severity group as indicated (g) in Otulin^∆hep and control mice at the age of 50–54 weeks. h Maximal tumour size in Otulin^∆hep mice, grouped by degree of pathology. Each data point represents the maximal tumour size in one mouse. Red bars indicate means ± SD. i Micrographs of H&E stained liver sections from Otulin^∆hep and control mice aged 50–54 weeks. (i) shows a control liver, (ii) shows HCC with abnormal macrotrabecular pattern, (iii) shows raised nuclear-cytoplasmic ratio, irregular nuclear outlines, and several mitotic figures (arrowheads), and (iv) shows a tumour with a steatohepatic appearance, with also enlarged and irregular nuclei. Micrographs are representative of four controls and 15 Otulin^∆hep mice. Tu tumour. NT non-tumour. b–d Data are presented as individual data points, each representing one mouse. Red bars indicate means. Data were analysed using an unpaired, two-sided Student’s t test. n.s., non-significant. See also Fig. S4.