Plko 1 egfp shrna lentiviral vectors

HUH7, HUH1 and MHCC97H cells were cultured in Dulbecco’s modified Eagle Medium (Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum (FBS), penicillin, streptomycin and L-glutamine (PSG). Hep3B cells were cultured in Minimum Essential Medium (Life Technologies) supplemented with 10% FBS, PSG, non-essential amino acids and sodium pyruvate. pLKO.1-PMPCB shRNA, pLKO.1-EpCAM shRNA and pLKO.1-eGFP shRNA lentiviral vectors were purchased from GE Healthcare (Lafayette, CO). R980-M15–663, a lentiviral vector with CAG promoter-driven firefly luciferase and eGFP expression was purchased from the Protein Expression Laboratory, Frederick National Laboratory for Cancer Research (Frederick, MD). Lentivirus particles were generated using Trans-Lentiviral shRNA Packaging System (GE Healthcare) or Lenti-vpak packaging kit (Origene, Rockville, MD). pCMV6-PMPCB-DDK plasmid was purchased from Origene. M50 Super 8× TOPFlash, M51 Super 8× FOPFlash and pCI-neo beta catenin S33Y plasmid were purchased from Addgene (Cambridge, MA). InSolution^™ Caspase Inhibitor VI (Z-VAD), GSK-3 Inhibitor IX (BIO) and GSK-3 Inhibitor IX Control (MeBIO) were purchased from Millipore (Billerica, MA). All cell lines were tested for mycoplasma and authenticated via STR analysis (August, 2015). Cells were passaged less than 15X after first thaw from liquid nitrogen.