Foetal bovine serum (fbs)

The human lung cancer cell lines A549, H358, and H460 and the lung epithelial cell line 16HBE were purchased from the Cell Bank at the Shanghai Institutes for Biological Sciences of the Chinese Academy of Sciences and were cultured in 1640 medium supplemented with 10% foetal bovine serum and 1% double antibiotics, as instructed by their provider. HEK293T and HeLa cells were cultured in DMEM and 10% foetal bovine serum (ExCell Bio, Shanghai, China) supplemented with 1% double antibiotics. All cells were maintained at 37°C with 5% CO2. The cells were treated with Importazole (ab146155) [27 (link)], Leptomycin B (ethanol solution, ab120501) or Bufexamac (S3023, Selleck, Shanghai, China) for the indicated times.

OSCC cell lines (SCC9, SCC15, SCC25) and Human Oral Keratinocytes (HOK) were obtained from Institute of Antibody Engineering, Southern Medical University. Cell lines HOK, SCC15 and SCC25 were seeded in DMEM (Gibco, Cat#11995500TB) and SCC9 in DMEM/F12 (Gibco, Cat#C11330500BT) containing 10% foetal bovine serum(FBS) (ExCell Bio, Inc) and incubated at 37°C with 5% CO₂.

A549 cells and MLE‐12 cells were preserved in our laboratory (National Collection of Authenticated Cell Cultures). A549‐shDNA‐PKcs and A549‐shNC cells were generated from A549 cells by transfecting the cells with lentiviruses carrying shRNA‐DPKcs‐U6/puromycin and shRNA‐NC vectors. The lentiviruses synthesised from GenePharma, and sequences shown as follow: shRNA‐DNA‐PKcs: 5ʹ‐GGG CGC TAA TCG TAC TGA A‐3ʹ; shRNA‐NC: 5ʹ‐TTC TCC GAA CGT GTC ACG T‐3ʹ. Cells were cultured at 37°C and 5% CO₂, supplemented with high‐glucose Dulbecco's Modified Eagle's Mudium (DMEM) containing 10% foetal bovine serum (Cat# FSP500, ExCell Bio) for nutrient, and 1% penicillin‒streptomycin solution (Cat# 2106030 M, Rongxia) was added to prevent cell contamination. Cells were irradiated with ⁶⁰Co γ‐rays at a dose of 6 Gy. For the VND3207 treatment group, the cells were first treated with 40 μM VND3207 (dissolved in dimethyl sulfoxide (DMSO)) for 2 h before IR.

The LX‐2 human hepatic stellate cell and HepG2 cells were obtained from the Institute of Biochemistry and Cell Biology (Chinese Academy of Sciences, Shanghai, China). The LX‐2 cells were cultured in DMEM (Solarbio technology) with 2% foetal bovine serum (FBS; Excell Bio).³⁰ We selected HepG2 as human hepatocyte cell line.³¹, ³² The HepG2 cells were cultured in DMEM (Solarbio Technology) with 10% foetal bovine serum (FBS; Excell Bio). All cells were cultured at 37°C in a 5% CO₂ incubator.

The normal oral epithelial cell line human oral keratinocytes (HOK) and OSCC cell lines SCC9, SCC25, and CAL27 were obtained from the Institute of Antibody Engineering, Southern Medical University (Guangzhou, China). Cell lines HOK, SCC25, and CAL27 were seeded in DMEM and SCC9 in DMEM/F12 containing 10% foetal bovine serum (FBS) (ExCell Bio Inc.) and incubated at 37°C with 5% CO₂.

Human embryonic kidney cells (HEK293T), normal liver cells (MIHA) and HCC cell lines (HepG2, Hep3B, Huh7, HCCLM3, PLC/PRF/5, and MHCC97L) were maintained in our laboratory [14 (link), 15 (link)]. Mycoplasma contamination was regularly detected using a Mycoplasma contamination detection kit (InvivoGen, San Diego, CA, USA). All cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco; Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% foetal bovine serum (FBS) (ExCell Bio, Shanghai, China) and incubated at 37 °C in 5% CO₂.