Generating Liver Cancer Tumors in Mice

Wild-type FVB/N mice were obtained from Charles River Laboratories (Wilmington, MA, USA) and the Rictor^fl/fl mice^{29 (link)} from the Jackson Laboratory (Sacramento, CA, USA). Hydrodynamic injection was performed as described previously.^{30 (link)} In brief, the plasmids encoding the genes of interest along with SB transposase in a ratio of 25:1 were diluted in 2 ml saline (0.9% NaCl), filtered through 0.22 μm filter, and injected into the lateral tail vein of the mice in 5–7 s. For the tumorigenesis models, 20 μg sgPten, 20 μg c-Met with 1.6 μg SB plasmid were delivered into FVB/N mouse liver (n=14). To determine the requirement of mTORC2 in sgPten/c-Met-driven liver carcinogenesis, high dose of Cre (60 μg) or pT3-EF1α (60 μg) was mixed with sgPten (20 μg), c-Met (20 μg) and SB (4 μg) and injected into Rictor^fl/fl mice (n=11 and 5, respectively). Mice were housed, fed and monitored in accordance with protocols approved by the Committee for Animal Research at the University of California, San Francisco.