Extended Data Fig. 7: Proliferation and YAP activation are not affected by increasing stiffness in low or high viscoelasticity hydrogels (Additional data to the main Fig. 5).
From: Matrix viscoelasticity promotes liver cancer progression in the pre-cirrhotic liver
a-e. Huh7 cells were encapsulated in low or high viscoelasticity IPN hydrogels with varying stiffness. (0.8-5 kPa). Cell proliferation and YAP activity were evaluated by Edu nuclear signal (a, scale bar, 100 μm) and active YAP immunofluorescence (b, scale bar, 50 μm), and quantification (c and d). e. mRNA expression of YAP target genes CTGF and Cyr6. (n = 4, for c-e, Error bars represent mean ± s.e.m. n numbers refer to independent experiments. One-way ANOVA with Tukey tests was used for correction of multiple comparisons. f-h. Huh7 cells were transfected with Tks5-mNeonGreen, and immunofluorescence microscopy was performed using an antibody against MT1-MMP (f), active integrin β1 (g), and p-MLC2 (h). i. Controls with no primary antibody for f-h. Cells were imaged by high resolution Airyscan microscopy (LSM980, Zeiss). Representative images, scale bar, 10 µm. j. Controls for the PLA assay in the main Fig. 5l (1° antibody only, 2° antibody only). Scale bar, 10 µm. Error bars represent mean ± s.e.m. n numbers refer to independent experiments. One-way ANOVA was used followed by Tukey’s multiple comparison test.
