Gbc sd

Human GBC cell lines were obtained as follows: GBC-SD and SGC-996, from the Shanghai Institutes for Biological Sciences (Shanghai, China), and OCUG-1 and NOZ were from the Japanese Collection of Research Bioresources JCRB cell Bank (Osaka, Japan). Human ovarian clear cell adenocarcinoma cell lines OVTOKO were also obtained from the JCRB Cell Bank. GBC-SD and OVTOKO cells were cultured in RPMI 1640 medium (Gibco, NY, USA), SGC-996 and OCUG-1 cells were cultured in Dulbecco’s Modified Eagle’s Medium (Gibco), and NOZ cells were cultured in William’s E medium (Gibco), with all media supplemented with 10% fetal bovine serum (Gibco), 10 units/ml penicillin, and 10 mg/ml streptomycin (1% P/S, Thermo Scientific HyClone, UT, USA). All cell lines were incubated at 37 °C in a humidified atmosphere containing 5% CO₂ and subcultured during the logarithmic phase. GBC cell lines and OVTOKO cells were authenticated by short tandem repeat profiling, as described previously. The short tandem repeat profiles are presented in Supplementary Figure S2.

The human gallbladder cancer cell lines GBC-SD, NOZ, SGC-996, OCUG, EHGB-1 and EHGB-2 were purchased from Shanghai Institute of Cell Biology, Chinese Academy of Sciences (CAS, Shanghai, China). The GBC-SD cells were grown in high-glucose DMEM (Gibco, Grand Island, NY, USA) supplemented with 10 % fetal bovine serum (Gibco) and 1 % penicillin-streptomycin(Hyclone, Logan, UT, USA). The NOZ cells were grown in William’s medium (Gibco) supplemented with 10 % fetal bovine serum (Gibco) and 1 % penicillin-streptomycin (Hyclone). The SGC cells were grown in 1640 medium (Gibco) supplemented with 10 % fetal bovine serum (Gibco) and 1 % penicillin-streptomycin (Hyclone), The OCUG, EHGB-1 and EHGB-2 cells were grown in high-glucose DMEM (Gibco) supplemented with 15 % fetal bovine serum (Gibco) and 1 % penicillin-streptomycin (Hyclone). All cells lines were maintained at 37 °C in a humidified atmosphere with 5 % CO₂. The A66 used in the in vitro and in vivo experiments was purchased from Selleck Chemicals (Houston, USA). Before conducting the CO-IP experiments, the GBC-SD and NOZ cells were starved for 12 h and 1 ng/mL EGF (Sangon Biotech, Shanghai, China) was added to the cell culture medium. The cells were maintained for 12 h to eliminate other confounding growth factor signals.

The human cell lines GBC-SD and SGC-996 were purchased from the Shanghai Cell Institute Country Cell Bank. GBC-SD cells were cultured in high-glucose DMEM (Gibco, USA) at 37°C in a 5% CO₂ incubator, while SGC-996 cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium. Both types of culture medium were supplemented with 10% fetal bovine serum (Gibco, USA), penicillin (100 U/mL) and streptomycin (100 U/mL) (Utah, HyClone, USA).

The human embryonic kidney 293T cells and human GBC cell line GBC-SD were purchased from the Chinese Academy of Life Sciences (Shanghai, China). Another GBC cell line NOZ was kindly gifted by Xinhua hospital (Shanghai, China).
GBC-SD and 293T cells were maintained in RPMI-1640 medium (GibcoBRL, Gaitherburg, MD, USA). NOZ cells were maintained in Willian’s E medium. All the cells were maintained in the medium supplemented with 1% antibiotics and 10% fetal bovine serum (GIBCO) at 37° C with 5% CO₂.

The human gallbladder cancer cell lines GBC-SD and NOZ were purchased from the Shanghai Cell Institute Country Cell Bank. GBC-SD cells were cultured in high-glucose Dulbecco’s modified eagle’s medium (DMEM) (Gibco, California, USA), and NOZ cells were cultured in William’s medium (Gibco) supplemented with 10% fetal bovine serum (Gibco), 100 μg/mL streptomycin, and 100 U/mL penicillin (Hyclone, Logan, UT) at 37°C in an atmosphere containing 5.0% CO₂. Cells were routinely grown in 100-mm plastic tissue culture dishes (Corning, New York, USA) and harvested using a trypsin-EDTA solution when they reached the logarithmic growth phase. Cells were maintained in these culture conditions for all experiments.