Hcc827

Parental H1975 (#CRL-5908) and HCC827 (#CRL-5908) cell lines were purchased from the ATCC (Manassas, VA, USA), and the PC9 cell line was obtained from the Varmus Laboratory (MSKCC, New York, NY, USA). All human lung adenocarcinoma cells were maintained in RPMI supplemented with 10% FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin. Jurkat cells procured from ATCC (#TIB-152) were cultured in RPMI with 10% FBS. Cells were authenticated by short tandem repeat (STR) profiling using the AmpFLSTR Identifiler kit (ThermoFisher, Waltham, MA, USA, #4322288) at the Protein Expression Laboratory (NCI, Frederick, Bethesda, MD, USA).

Lung adenocarcinoma cell line HCC827, lung carcinoma cell line NCI-H358 (H358) and mammary ductal carcinoma cell line T-47D were purchased from ATCC (Manassas, VA) and grown in RPMI 1640 culture medium containing 10 % FBS, 100 units/ml of penicillin, and 100 μg/ml streptomycin in a humidified air (5 % CO₂ atmosphere) at 37 °C. Each of three lentivirus claudin-7 shRNA vectors (sequences #1: 5′-TTCCAAGGAGTATGTGTGA-3′; #2: 5′-GGCTATGGGAGTGTCTAGA-3′; #3: 5′-TCCCTACCAACATTAAGTA-3′) or one off-target shRNA control vector were transfected into HCC827, H358 or T-47D cells. The cells infected with #2 lentivirus claudin-7 shRNA vector are designated as KD cells, and the cells infected with #3 shRNA vector are designated as KD2 cells. All lentivirus vectors contain a GFP expression sequence. After 48 h incubation, transfected cells were selected by 1 μg/ml puromycin.
The pcDNA3.1 myc or pcDNA3.1-claudin-7-myc (mouse claudin-7 cDNA) was transfected into HCC827 claudin-7 KD cells using Amaxa Nucleofector device (Amaxa Biosystems, Cologne, Germany). Geneticin (G418) was added to select the transfected cells.

A549, PC9, HCC827 and HSAEC1-KT cells were obtained from ATCC (Manassas, VA, USA) and CP-resistant cells were generated with the procedures as described previously.^{34 (link), 35 (link)} The human bronchial epithelial cell line 16HBE was a kind gift from Sino-French Hoffman Institute, Guangzhou Medical University, Guangdong, China. All cells were cultured based on the guidance from the manufacturer. CP resistance was maintained at 5 μM of CP during culture. All cells were grown as monolayer cultures and maintained in a humidified atmosphere of 5% CO₂ in air at 37 °C. All the cells have been authenticated before use.

The human lung adenocarcinoma cell lines (A549 and HCC827) were obtained from ATCC. The cells were cultured in an humidified atmosphere with 95%air and 5%CO₂ at 37°C,and supplemented with RPMI-1640 medium (Gibco, ThermoFisher, Shanghai, China) with 10% heat-inactivated fetal bovine serum, streptomycin (100 U/mL) and penicillin (100 U/mL). A549 and HCC827 cells were split every 2–3 days at a concentration of 1.5^*105 and cells/ml.

All cell lines, HT-29, SK-CO-1, NCI-H747, and HCC-827, were purchased from ATCC (Manassas, VA, USA). ATCC authenticates all its cell lines by short tandem repeat profiling, cell morphology monitoring, karyotyping, and cytochrome C oxidase I testing. The cells were grown in ATCC-recommended media, containing 10% FBS and 1% Penicillin-streptomycin (Thermo Fisher Scientific, Pittsburgh, PA, USA). All cells were cultured at 37 °C in humid atmosphere containing 5% CO₂. Kinase inhibitors were purchased from Selleckchem (Munich, Germany), LC Laboratories (Woburn, MA, USA) or AdooQ Bioscience (Irvine, CA, USA).

A549 (CCL-185), NCI-H460 (HTB-177), NCI-H1975 (CRL-5908), and HCC827 (CRL-2868) cells were purchased from ATCC and cultured in RPMI supplemented with 10% (v/v) FBS, glutamine (4 mM), and penicillin–streptomycin (100 U mL⁻¹). The cell lines were tested for mycoplasma contamination but were not authenticated. All cell lines were transduced with a plasmid that expresses nuclear-localized blue-fluorescent protein (FU-EBFP2-H2B-W; obtained from the UCLA Molecular Screening Shared Resource (MSSR)).