Fig. 2: Mice on a HiAD develop more transformed foci, and exhibit AGE-dependent higher viscoelasticity.
From: Matrix viscoelasticity promotes liver cancer progression in the pre-cirrhotic liver
a, Schematic of the NASH-related HCC model (early timepoint). Mice were fed for 7 weeks with chow, FFD or HiAD. HDI was performed using vectors expressing human MET (pT3-EF5a-hMET) and sleeping beauty (SB) transposase combined with a vector expressing either wild-type (pT3-EF5a-β-catenin-MYC, control) or mutant (pT3-EF5a-S45Y-β-catenin-MYC) human β-catenin. Then, 4 weeks after HDI, the mice were euthanized. b, Immunohistochemistry on sequential slides for the co-localization of GS- and MYC-tag-positive foci (circles, more than 20 cells are considered to form a focus). Scattered positive cells denote transduced cells (arrows). GS at the baseline marks pericentral cells. c, Quantification of GS- and MYC-tag-positive foci 4 weeks after HDI. n = 5 each. d, Schematic of the NASH-related HCC model combined with AGE-lowering approaches; mice were euthanized 7 weeks after HDI. e, Immunohistochemistry on sequential slides showing the co-localization of GS- and MYC-tag-positive foci (circles). Scattered positive cells denote transduced cells (arrows). f, The survival rates of mice after HDI. n = 7 (chow), n = 7 (FFD), n = 11 (HiAD + vehicle), n = 7 (HiAD + PM) and n = 7 (HiAD + ALT) mice. g, Quantification of transformed foci 7 weeks after HDI. Left to right, n = 8, 8, 6, 7 and 7, 20 areas per mouse. h, Schematic of the NASH-related HCC model combined with AAV8-mediated AGER1 induction before HDI. i,j, Liver AGER1 (also known as Ddost) mRNA expression (i) and AGE levels (j) were assessed. n = 5 each. k–m, Rheometry data of studies on stiffness (k) and viscoelasticity (l, m) in AGER1-reconstituted mice. n = 5 each. n,o, Quantification of transformed foci (n = 5 each, 20 areas per mouse) (n) and GS and MYC immunohistochemistry analysis on sequential slides (o). Data are mean ± s.e.m. n values refer to individual mice. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple-comparison test. For b, e and o, scale bars, 300 μm. ALT, alagebrium. The diagrams in a, d and h were created using BioRender.
