Extended Data Fig. 10: PTB ___domain deleted (dd) TNS-1 did not colocalize with integrin, and cells exhibited lower proliferation, YAP activity, and reduced formation of invadopodia-like structures in high viscoelasticity hydrogels.
From: Matrix viscoelasticity promotes liver cancer progression in the pre-cirrhotic liver
a. Schematics of the PTB ___domain deleted (dd) TNS1 construct. Deleting this ___domain prevents binding to integrins however the actin-binding ___domain remains intact. b, c. Proximity ligation assays (b) to assess integrin β1 and TNS1 binding (green signal) in empty vector (NC), full length TNS1, and dd-TNS1 transfected cells (red, tdTomato). In full length TNS1 tomato, PLA signals colocalized whereas no colocalization was seen in dd-TNS1 transfected cells or in low viscoelasticity matrix. PLA positive dots were quantified from 30 cells in 5 gels, each group (c, n = 5 each). d-k. Cell proliferation was evaluated by Edu (d, Scale bar, 100 μm) and quantification (h). YAP activity was analysed by using active YAP immunofluorescence (e, Scale bar, 50 μm.), quantification (i), and YAP-regulated target gene mRNA expression (k). Cell circularity was evaluated by F-actin signal (f, Scale bar, 50 μm, and) and quantification (j, ImageJ). Formation of invadopodia-like structures was analysed by the TKS5 signal (g, Scale bar, 10 μm, n = 5 each. Error bars represent mean ± s.e.m. n numbers refer to independent experiments. One-way ANOVA test was used followed by Tukey’s multiple comparison.
