Ha22t

HCC cell lines, SKHEP1 (RRID: CVCL_0525), HepG2 (RRID: CVCL_0027), SNU387 ((RRID: CVCL_0250), and Huh7 (RRID: CVCL_0336) were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan), while the cell lines Hep3B (RRID: CVCL_0326), HA22T (RRID: CVCL_7046), and PLC5 (RRID: CVCL_0485) were obtained from American Type Culture Collection (Manassas, VA, USA). All the cell lines were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Thermo Fisher Scientific, Grand Island, NY, USA), which was supplemented with 10% Fetal Bovine Serum (FBS) (Thermo Fisher Scientific), 100 IU/mL penicillin, and 100 μg/mL streptomycin (Thermo Fisher Scientific). The culturing of the cells was performed in a humidified tissue culture incubator with a 5% CO₂ atmosphere at 37 °C. All human cell lines have been authenticated using short tandem repeat profiling within three years. All experiments were performed with mycoplasma-free cells.

HCC cell lines, SKHEP1 (RRID: CVCL_0525), HepG2 (RRID: CVCL_0027), SNU387 ((RRID: CVCL_0250), and Huh7 (RRID: CVCL_0336) were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan), while the cell lines Hep3B (RRID: CVCL_0326), HA22T (RRID: CVCL_7046), and PLC5 (RRID: CVCL_0485) were obtained from American Type Culture Collection (Manassas, VA, USA). All the cell lines were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Thermo Fisher Scientific, Grand Island, NY, USA), which was supplemented with 10% Fetal Bovine Serum (FBS) (Thermo Fisher Scientific), 100 IU/mL penicillin, and 100 μg/mL streptomycin (Thermo Fisher Scientific). The culturing of the cells was performed in a humidified tissue culture incubator with a 5% CO₂ atmosphere at 37 °C. All human cell lines have been authenticated using short tandem repeat profiling within three years. All experiments were performed with mycoplasma-free cells.

We obtained three human HCC cell lines from the American Type Culture Collection (ATCC, Rockville, MD): HepG2, HA22T, Hep3B, and TONG. They were all grown in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% (w/v) fetal bovine serum, penicillin at 100 U/mL, streptomycin at 100 μg/mL, and L-glutamine at 2 mmol/L (all from Invitrogen, Carlsbad, CA) at 37°C in an atmosphere of 5% (v/v) CO₂ in air.

Human HCC cell lines including HepG2, HA22T, MHCC-LM3, Huh7, JHH-7, and HLF were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The human immortalized liver cell line of L02 was bought from the China Center for Type Culture Collection (CCTCC, China). Dulbecco’s modified Eagle’s medium (DMEM, Gibco, USA) was used to culture HepG2, MHCC-LM3, Huh7, JHH-7, and HLF cells. Meanwhile, L02 and HA22T cells were cultured in RPMI-1640 medium (Gibco, USA). All mediums used in the study contained 10% fetal bovine serum (FBS, Gibco, USA) and 1% penicillin/streptomycin (Beyotime, China). Then, the cells were incubated at a constant temperature and humidity incubator at 37 °C with 5% CO₂.
Small interfering RNA (siRNA) targeting TBK1 designed by Hanbio Biotechnology, Shanghai, China, and exogenous TBK1 plasmid purchased from Bochu Biotechnology, Changsha, China, were transfected into cells using Lipofectamine 3000. TBK1-siRNA target sequences (5′–3′) #1: AAGGUACUGGCAAUUCUGCTT. #2: AUUGUUCCCUGAGAACUGGTT.

Male BALB/c nu/nu mice were obtained from the National Laboratory of Animal Breeding and Research Center (Taipei, Taiwan) and housed according to the protocols of the Animal Center of the Kaohsiung Medical University (Kaohsiung, Taiwan). Spz1 transgenic mice were generated by Dr. Hung Li (Academia Sinica, Taipei, Taiwan) [11 (link), 12 (link)]. Human hepatoma cell lines (Hep G2, Hep 3B, SK-Hep1, Huh 7, PLC/PRF/5, and HA 22T) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA) and maintained according to ATCC protocols. The study was conducted with approval (IACUC-104181) from the ethics committee of Kaohsiung Medical University.