LN229, A431, Chinese hamster ovary (CHO)-K1, HEK-293T, MCF-10A, Met-5A, and P3U1 cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA). HSC-2 and HSC-3 were obtained from the Japanese Collection of Research Bioresources (JCRB) Cell Bank (Osaka, Japan). LN229/EGFR and CHO/EGFR were produced by transfecting pCAG/PA-EGFR-RAP-MAP(12 (link)) into LN229 and CHO-K1 cells using Neon transfection system (Thermo Fisher Scientific, Inc., Waltham, MA) and Lipofectamine LTX (Thermo Fisher Scientific, Inc.), respectively. A few days after transfection, PA tag-positive cells(13 (link)) were sorted using a cell sorter (SH800; Sony Corp., Tokyo, Japan).
CHO-K1, CHO/EGFR, and P3U1 cell lines were cultured in RPMI 1640 medium (Nacalai Tesque, Inc., Kyoto, Japan), and LN229, LN229/EGFR, A431, HSC-2, HSC-3, HEK-293T, MCF-10A, and Met-5A cell lines were cultured in Dulbecco's modified Eagle's medium (Nacalai Tesque, Inc.), supplemented with 10% heat-inactivated fetal bovine serum (Thermo Fisher Scientific, Inc.), 100 units/mL of penicillin, 100 μg/mL of streptomycin, and 25 μg/mL of amphotericin B (Nacalai Tesque, Inc.) at 37°C in a humidified atmosphere containing 5% CO2 and 95% air.
CHO-K1, CHO/EGFR, and P3U1 cell lines were cultured in RPMI 1640 medium (Nacalai Tesque, Inc., Kyoto, Japan), and LN229, LN229/EGFR, A431, HSC-2, HSC-3, HEK-293T, MCF-10A, and Met-5A cell lines were cultured in Dulbecco's modified Eagle's medium (Nacalai Tesque, Inc.), supplemented with 10% heat-inactivated fetal bovine serum (Thermo Fisher Scientific, Inc.), 100 units/mL of penicillin, 100 μg/mL of streptomycin, and 25 μg/mL of amphotericin B (Nacalai Tesque, Inc.) at 37°C in a humidified atmosphere containing 5% CO2 and 95% air.
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