Extended Data Fig. 6: YAP pathways are induced by matrix viscoelasticity.
From: Matrix viscoelasticity promotes liver cancer progression in the pre-cirrhotic liver
a. Analyses of bulk RNA-seq data from mice fed chow, FFD, and HiAD diets. HiAD-fed mice were treated by PM or vehicle, and a group of mice with RAGE hepatocyte depletion (RAGEHepKO) were studied. Genes between FFD and HiAD groups with Log2 fold change (Log2FC) and p-value less than 0.05 (Fisher’s exact test) were considered differentially expressed. KEGG analyses from differentially expressed genes show enrichment in Hippo signalling pathway. No HD injection was done in these experiments. b, c. Representative images (b) and quantification (c) of active nuclear YAP signal in mouse livers, using an antibody against the active, non-phosphorylated YAP. (n = 5 mice/group, 4 random ×20 fields/sample; data are presented as the percentage of active YAP/area/×20 field). d. YAP targets CTGF and Cyr61 were induced in mice on HiAD, but not after PM or in RAGEHepKO on HiAD. RT-qPCR, n = 6 each. e. Collagen was imaged using SHG (green), active YAP (red) and α-SMA positive stellate cells (blue, arrows) by immunofluorescence. There were no hepatocytes with active YAP observed in the close proximity of collagen bundles (using an antibody against the non-phosphorylated active YAP). Scale bar 50 µm. f. HE images corresponding to sequential slides with myc-tag/GS immunohistochemistry in the main Fig. 4c. g. Nuclear and cytoplasmic YAP was assessed using antibodies against active, non-phosphorylated and inactive phosphorylated YAP in western blots (cytoplasmic and nuclear fractions), in low and high viscoelasticity hydrogels (representative of 3 different experiments). Error bars represent mean ± s.e.m. n numbers refer to individual mice. One-way ANOVA was used followed by Tukey’s multiple comparison test.
