Ha and β actin

Doxycycline hyclate (DOX, Sigma); ICG-001 (Selleckchem); fetal bovine serum, DMEM and DMEM/F12 medium (Gibco); fibroblast growth factor (FGF) and epidermal growth factor (EGF) (PeproTech); Trizol reagent (TAKARA); protease and phosphatase inhibitor cocktail (Roche); Lipofectamine 3000 and B27 (Invitrogen); dual-specific luciferase assay kit (Promega); Cell Counting Kit-8 (CCK8, Dojindo Molecular Technologies); Western blotting substrate (Millipore); Cell Signaling Senescence β-Galactosidase Staining Kit (CST); silver staining Rapid silver staining kit's (Beyotime); BALB/c nude mice (Beijing Vital River Laboratory Animal Technology); antibody against GATA4, Lamin B1, P21, FLAG, P15 and c-MYC (Abcam); HA and β-actin (Sigma); β-catenin, LEF1, TCF1, P14, P27, P53, p14/ARF, Caspase-9 and Caspase-3 (CST), P16/Ink4a (Epitomics); Cytokeratin (AE1/AE3) antibody (Kit-0009, MXB Biotechnologies) were purchased from the indicated manufacturers. SNU-387, SNU-449, PLC, NeHepLxHT, SK-Hep1, HepG2, HUH7 and HEK293 cells were obtained from ATCC.

Antibodies against the following proteins were obtained from the indicated suppliers: Na⁺/K⁺ ATPase, GFP and tubulin (Santa Cruz Biotechnology); c‐Jun, p‐c‐Jun, ERK, p‐ERK, c‐MET, p‐c‐MET and GST (Cell Signaling Technology); V5, Myc (MBL); HA and β‐actin (Sigma); KITENIN (Atlas); His and RACK1, LC3, p62, Myo10, and eukaryotic elongation factor 2 (eEF2) (Abcam). They were used with appropriate secondary antibodies (Thermo). For transient transfection analyses, Caco2, HCT116 and 293T cells were transfected with various plasmids and harvested for immunoblot analysis 48 h after transfection. For most assays using stable cell lines, mixed polyclonal cells were used to exclude clonal variation. Cellular proteins were separated, transferred and immunoblotted as previously described.¹⁸ Cell lysates from Caco2, HCT116 and 293T cells were used for immunoprecipitation experiments as previously described.¹⁹