Pcmv6 ddk myc empty vector

The human normal bronchial epithelial (HBE), and the A549, PC9 cancer cell lines were obtained from the American Type Culture Collection (ATCC; Manassas, VA). The H1299 and H460 cancer cell lines were obtained from the Shanghai Cell Bank (Shanghai, China). The LK2 cell line was a gift from Dr. Hiroshi Kijima (Department of Pathology and Bioscience, Hirosaki University Graduate School of Medicine, Hirosaki, Japan). All the cell lines were cultured in RPMI 1640 (Invitrogen, Carlsbad, CA, USA) or DMEM (Invitrogen) containing 10% calf serum at 37°C and 5% CO2. The pCMV6-ddk-myc empty vector, pCMV6-ddk-myc-FAM110B plasmid (RC206875), and the negative control (NC)-siRNA (SR30004) were all purchased from Origene (Rockville, MD, USA). The FAM110B-siRNA (sc-77299) was purchased from Santa (Santa Cruz Biotechnology, Inc. CA, USA) which composed of a pool of two target-specific 19–25 nt siRNAs. Lipofectamine 3000 transfection reagent (Invitrogen) was used for cell transfections. After transfection, cells were incubated for 48 hours and analyzed using Western blot. For specific inhibition of Wnt/β-catenin signaling, a Wnt/β-catenin signaling inhibitor, XAV-939, was incorporated into the medium of the cultured cells at a concentration of 30nM for 24 hours at 37°C (S1180, Selleck Chemicals, Houston, TX, USA).