Hcc2279

The H358, H460, NCIH23, SKLU-1, SW960, A427, H727 (KRAS^Mut), H1650 (KRAS and EGFR double mutant), H1975, HCC827, HCC2279 (EGFR^Mut), HCC1666, H322M, H522, Calu-3, H1299 (KRAS and EGFR wild type), BEAS-2B (nontumorigenic lung epithelial cells), CCD-18Lu (nontumorigenic lung fibroblasts), and human embryonic kidney (HEK) 293 T cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA). The GFP-positive KRAS^G12C mutant stable cell lines H1299 KRAS^G12C_K104, H1299 KRAS^G12C_K104R, and H1299^H363Y were generated by stably cotransfecting H1299 KRAS^WT cells. The cells were cultured in either HyClone RPMI-1640 medium (Thermo Fisher Scientific, Waltham, MA, USA) or Dulbecco’s modified Eagle’s medium (Thermo Fisher Scientific, Waltham, MA, USA), each containing 10% FBS and 1% penicillin and streptomycin. Cells were maintained in a 37 °C humidified incubator with 5% CO₂ in air. All cell lines were authenticated by short tandem repeat (STR) analysis and were free of mycoplasma contamination.

A549, Calu-6, NCI-H1299, NCI-H1703, NCI-H1975, NCI-H23, NCI-H460, HCC2279, NCI-H522, A427, Calu-3, NCI-H358, HCC827 and BEAS2B were obtained from the American Type Culture Collection (Rockville, MD, USA). HCC95 and HCC1588 were obtained from the Korean Cell Line Bank (Seoul, Korea), and Normal human bronchial epithelial (NHBE) purchased from Lonza. A549, Calu-6, NCI-H1299, NCI-H1703, NCI-H1975, NCI-H23, NCI-H460, HCC2279, NCI-H522, NCI-H358, HCC827, HCC95, HCC1588, and BEAS2B were cultured with RPMI1640. Calu-3 and A427 were cultured with Dulbecco’s modified eagle medium (DMEM). NHBE was cultured with bronchial epithelial cell growth medium (BEGM) BulletKit (Lonza, Walkersville, MD, USA). All cell lines were maintained in media supplemented with 10% fetal bovine serum (FBS) and 1× penicillin-streptomycin, and cultured under standard conditions at 37 °C in a humidified atmosphere of 95% air and 5% CO₂. Stock solutions of the Wee1 inhibitor MK-1775 (Selleck Chemicals, Houston, TX, USA) were dissolved in dimethyl sulfoxide (DMSO) and added to the media at the indicated concentrations (100 nM). Doxycycline (D9891) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Matrigel was purchased from Corning Inc (Corning, NY, USA).