Extended Data Fig. 3: Collagen networks in patients, mouse models and 3D hydrogels.
From: Matrix viscoelasticity promotes liver cancer progression in the pre-cirrhotic liver
a. Collagen fibres in human liver samples (healthy, NASH, NASH+ T2DM) were analysed by second harmonic generation (SHG) microscopy. Areas with bundling is depicted in the NASH + T2DM group (white arrow). Red arrow points to lipid droplets. Maximum intensity projection Z-stack for section thickness of 30 μm (Representative images from three individual subjects. Scale bar, 200 µm). b. In mouse livers images depict a more bundled appearance of collagen network (white arrow) in the HiAD+vehicle group whereas more organized fibres following PM, ALT treatment or after AGER1 reconstitution. Maximum intensity projection Z-stack for section thickness of 30 μm (Representative images from three individual mice. Scale bar, 200 µm). c. Local patches with higher Young’s moduli could be observed in collagen hydrogels (red arrows). d. The ranges of elastic moduli is depicted in collagen or collagen+AGEs hydrogels. e. Mapping and distribution of hysteresis areas (viscoelasticity). f. Higher frequencies of increased viscoelasticity in collagen+AGEs gels. n = 5 gels/each group, 3 representative areas/each gel. For all maps, x’s indicate regions where AFM indentation curves could not be reliably analysed. Scale bar, 20 µm. g-j. To analyse the potential effect of ALT on the matrix, collagen gels +/− ALT-711 were studied by SHG (g, Scale bar, 100 µm). Gels from g were loaded for rheometry to assess stiffness (h, storage modulus) and viscoelasticity (i, loss tangent; j, τ1/2), (n = 4 each). Error bars represent mean ± s.e.m. n numbers refer to independent experiments. Two-tailed, unpaired t-tests for statistical analysis. ns, not significant.
