Ln229

P3X63Ag8U.1 (P3U1), Chinese hamster ovary (CHO)-K1, LN229, and NMuMG (a mouse mammary gland epithelial cell) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA). Lewis lung carcinoma was obtained from the Japanese Collection of Research Bioresources (JCRB; Osaka, Japan).
The pCAG-zeo_ssnPA-mEGFR and pCAG-zeo_MAP-mEGFR plasmids were transfected into LN229 and CHO-K1 cells, respectively. The stable transfectants were generated as described previously [40 (link),41 (link)].
CHO-K1, mEGFR-overexpressed CHO-K1 (CHO/mEGFR), Lewis lung carcinoma, and P3U1 were cultured in an RPMI-1640 medium (Nacalai Tesque Inc., Kyoto, Japan), with 10% fetal bovine serum (FBS; Thermo Fisher Scientific Inc., Waltham, MA, USA). A cocktail of 100 units/mL of penicillin, 100 μg/mL of streptomycin, and 0.25 μg/mL of amphotericin B (Nacalai Tesque Inc.) was added to the medium. LN229, mEGFR-overexpressed LN229 (LN229/mEGFR), and NMuMG were cultured in DMEM (Nacalai Tesque Inc.), supplemented as indicated above. For NMuMG cells, 10 μg/mL of insulin (Sigma-Aldrich Corp., St. Louis, MO, USA) was further added. All cells were cultured using a humidified incubator at 37 °C, in an atmosphere of 5% CO₂ and 95% air.

COLO205 (a human colorectal cancer cell line) was obtained from the Cell Resource Center for Biomedical Research Institute of Development, Aging and Cancer at Tohoku University (Sendai, Japan). HSC-3 (a human OSCC cell line) and LN229 (a human glioblastoma cell line) were obtained from the Japanese Collection of Research Bioresources (Osaka, Japan). P3X63Ag8U.1 (P3U1: a mouse multiple myeloma) and CHO-K1 cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). HSC-3 and LN229 were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Nacalai Tesque, Inc., Kyoto, Japan), supplemented with 10% (v/v) heat-inactivated fetal bovine serum (FBS; Thermo Fisher Scientific, Inc., Waltham, MA, USA), 100 U/mL of penicillin, 100 μg/mL streptomycin, and 0.25 μg/mL amphotericin B. CHO-K1, COLO205, and P3U1 were cultured in Roswell Park Memorial Institute (RPMI)-1640 medium (Nacalai Tesque, Inc.) supplemented with 10% FBS and antibiotics, as indicated above. All the cells were grown in a humidified incubator at 37 °C with 5% CO₂.