Penicillin streptomycin solution

HeLa cells, a human cervical carcinoma cell line, were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). HeLa/Fucci cells, a subline of the HeLa cell line that expresses a cell cycle marker Fucci [23 (link)], and Hepa 1–6 cells, a mouse hepatoma-derived cell line, were obtained from the RIKEN Bioresource Center (Tsukuba, Japan). All of the cell types were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM; Fujifilm Wako Pure Chemical, Osaka, Japan; 044-29765) supplemented with 10% fetal bovine serum (FBS; Biowest, Riverside, MO, USA) and a penicillin-streptomycin solution (Fujifilm Wako Pure Chemical; 168-23191) at 37 °C in a 5% CO₂ incubator. Medium that was free of both Met and cystine was prepared by using DMEM/high glucose/no glutamine/ no methionine/no cystine (Thermo Fisher Scientific, Waltham, MA, USA; 21013-024) supplemented with 10% FBS, a penicillin-streptomycin solution, 4 mM L-glutamine (Fujifilm Wako Pure Chemical), and 1 mM sodium pyruvate (Fujifilm Wako Pure Chemical). For cystine deprivation, the above medium was further supplemented with 0.2 mM L-methionine (PEPTIDE INSTITUTE, Osaka, Japan). Erastin (Item No. 17754), SAM (Item No. 16376), SAH (Item No. 13603), and Hcy (Item No. 30852) were purchased from Cayman Chemical (Ann Arbor, MI, USA).

The A673 cell line was purchased from the European Collection of Authenticated Cell Cultures (ECACC) and cultured in Dulbecco’s Modified Eagle Medium (DMEM, Cat. No. 044–29765, Fujifilm-Wako chemical) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 1x Penicillin-Streptomycin Solution (Cat. No. 168–23191, Fujifilm-Wako chemical). The Seki cell line was established by Nojima et al. [20 (link)], purchased from the Cell Resource Center for Biomedical Research, Institute of Development, Aging and Cancer, Tohoku University (Cat. No. TKG 0725, Miyagi, Japan), and cultured in RPMI-1640 (Cat. No. 189–02025, Fujifilm-Wako chemical) with 10% FBS and 1x Penicillin-Streptomycin Solution. The NCR-EW2 cell line and SAOS-2 cell line (ATCC HTB-85) were cultured in RPMI-1640 with 10% FBS and 1x Penicillin-Streptomycin Solution. Human Embryonic Kidney cells 293 (HEK293) cells and hTERT RPE-1 (ATCC CRL-400), Lenti-X^™ 293T cell line (Cat. No. 632180, Takara-bio), and U-2 OS cell line were cultured in DMEM with 10% FBS and 1x Penicillin-Streptomycin Solution. Seki, NCR-EW2, and Lenti-X293T cells were spread onto a 0.1% gelatin-coated dish.

Authenticated cell lines, COS-7 (Cell #. JCRB9127) and HEK293 (JCRB9068), were obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan). The cells were cultured in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS; Equitech-Bio, Kerrville, TX, USA) and 1% penicillin/streptomycin solution (Fujifilm Wako Pure Chemical Corporation, Osaka, Japan). The cells were maintained according to our previous studies [35 (link),36 (link)]. For transfection, cells were cultured on appropriate culture plates or dishes, with lipofection of plasmid vectors using TransFast Transfection Reagents (Promega) according to the manufacturer’s protocols.

MCF-7 human breast cancer cells provided by the American Type Culture Collection (ATCC) were cultured in Dulbecco’s modified Eagle’s medium (Nissui Pharmaceutical) supplemented with 10% of foetal bovine serum (Sigma-Aldrich) and 1% of a penicillin/streptomycin solution (Wako Pure Chemical Industries).

Mouse B16 melanoma 4A5 cells [38 (link)] were obtained from RIKEN Cell Bank (Ibaraki, Japan) and were cultured in α-minimum essential medium (Nacalai Tesque, Tokyo, Japan) with 10% fetal bovine serum (Biowest SAS, Nuaille, France) and 1% penicillin–streptomycin solution (Wako Pure Chemicals, Osaka, Japan) at 37 °C in 5% CO₂.