H1975

Human lung epithelial cell line (H1975) and liver cell line (HepG2) were obtained from the American Type Culture Collection (ATCC^®); H1975 cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium and HepG2 cells were cultured in high-glucose Dulbecco’s modified Eagle medium (DMEM) (see Tables S2 and S4 of the SI), both supplemented with 10% FBS and 1% penicillin–streptomycin in a 5% CO₂ atmosphere at 37 °C. CB was heated at 200 °C for 120 min to sterilize, then resuspended by sonication in complete RPMI and DMEM medium with 2% Tween 80 at concentrations of 50, 100, and 200 μg mL⁻¹. The working solutions were freshly diluted 40 times in complete RPMI and DMEM media to concentrations of 1.25, 2.5, and 5 μg mL⁻¹. H1975 and HepG2 cells were treated with CB for 48 h. RPMI and DMEM medium with 0.05% Tween 80 was used as control.

Human lung cancer cell lines H1975 (ATCC^® CRL-5905^TM), H838 (ATCC^® CRL-5844^TM), and H226B were purchased from American Type Culture Collection (Manassas, VA). H1975 cells, which carry the EGFR L858R/T790M mutation, were cultured in RPMI-1640 containing high glucose, L-glutamine, and HEPES (ATCC #30–2001; American Type Culture Collection, Manassas, VA), supplemented with 10% fetal bovine serum, at 37°C in 5% carbon dioxide. H838 cells, which carry the EGFR L858R mutation, were cultured in RPMI-1640 supplemented with 10% fetal bovine serum at 37°C in 5% carbon dioxide. H226B cells, which are a human squamous cell lung cancer cell line and do not carry any EGFR mutations, were cultured in RPMI-1640 supplemented with 10% fetal bovine serum at 37°C in 5% carbon dioxide.

Lung cancer cell lines (A549, H460, H1299, H1975, H358), human bronchial epithelial cell line BEAS-2B, and mouse lewis lung cancer cells (LLC) were all purchased from the ATCC (USA). YFP-53BP1-HT1080 cells were a kind gift from Department of Radiation Oncology, UT Southwestern Medical Center. M059K and M059J were purchased from KeyGENE BioTech Co., China. A549, H460, H1975, H358 and HT1080 were maintained in DMEM with 10 % fetal bovine serum at 37℃ in a 5 % CO₂ humidified chamber. BEAS-2B, H1299 and LLC cells was maintained in RMPI 1640 medium with the same supplement. Cells were treated with ionizing radiation, Etoposide, CPT, H₂O₂, 4NQO or UV to induce different types of DNA damages. Specific inhibitors for ATM, ATR and DNA-PKcs, known as KU55933, VE821 and NU7441, were purchased from Sellecks Biotech. For laser irradiation, a 365-nm pulsed nitrogen laser was directly coupled to the fluorescence path of the microscope (Axiovert 200 M; Carl Zeiss).

Human cell lines A549 (EGFR wild-type) and H1299 (EGFR wild-type) were purchased from ATCC, while H1975 (EGFR L858R and T790M mutations), HCC827 (EGFR ΔE746-E750) and HCC4006 (EGFR ΔL747-E749) cell lines were a gift of Dr. Ming Tsao (Princess Margaret Hospital, Toronto, ON). A549 cells were maintained in DMEM (Mediatech, Manassas, VA) supplemented with 10% fetal bovine serum (FBS; Medicorp, Montreal, QC), while H1299, H1975, HCC827 and HCC4006 cells were maintained in RPMI-1640 (HyClone Laboratories, Logan, UT) with 10% FBS. All cell lines were grown at 37°C and 5% CO₂. The FAK inhibitor PF-573,228 (PF-228) was purchased from Tocris Bioscience (Bristol, UK) and dissolved in DMSO. Erlotinib and the FAK inhibitor PF-562,271 (PF-271) were purchased from Shanghai Biochempartner Co. Ltd. (Shanghai, China) and dissolved in DMSO for in vitro cell culture work.