Extended Data Fig. 12: TNS-1 knockdown decreases the formation of transformed foci in mice on HiAD, after HDI.
From: Matrix viscoelasticity promotes liver cancer progression in the pre-cirrhotic liver
a. Schematic presentation of the in vivo targeting of TNS1 by CRISPR/Cas9 in conjunction with hydrodynamic injection (HDI). Mice were fed chow or HiAD for 7w, then hydrodynamically injected with pT3-EF5a-hMet and the pT3-354 EF5a-S45Y-β-catenin-myc (mutant β-catenin), with the sleeping beauty (SB) transposase, as well as the CRISPR-Cas9-based vector either with sgRNAs targeting mouse TNS1 (pX333-sgTNS1) or empty vector (sgNC). Mice were sacrificed 7 weeks following injection. b. TNS1 expression in the liver was analysed by RT-qPCR (n = 6 each). c, d. H&E images, GS/myc immunohistochemistry on consecutive slides depict colocalization (c). Quantification (d) of GS/myc positive foci. Scale bar, 300 μm (n = 6 each). e. The expression of YAP targets CTGF and Cyr 61 was analysed by RT-qPCR (n = 6 each). Error bars represent mean ± s.e.m. n numbers refer to individual mice. One-way ANOVA test was used followed by Tukey’s multiple comparison.
