Sodium pyruvate solution

The estrogen receptor (ER)-positive human breast cancer line MCF-7 (RCB1904) and the ER-negative human breast cancer cell line MDA-MB-453 (RCB1192) were purchased from the RIKEN cell bank. MCF-7 cells were cultured at 37 °C with 5% CO₂ in E-MEM with L-glutamine and phenol red (Wako Pure Chemical Industries, Ltd., Osaka, Japan) containing 10% fetal bovine serum (GE Healthcare Life Sciences, Inc., Logan, UT, USA), 1% penicillin/streptomycin (Wako Pure Chemical Industries, Ltd.), 1.0 mM sodium pyruvate solution (Wako Pure Chemical Industries, Ltd.), and 1% MEM non-essential amino acid solution (Thermo Fisher Scientific, Inc., Waltham, MA, USA). MDA-MB-453 cells were cultured at 37 °C without CO₂ in Leibovitz′s L-15 Medium with L-glutamine, phenol red, and sodium pyruvate (Wako Pure Chemical Industries, Ltd.) containing 10% fetal bovine serum (GE Healthcare Life Sciences, Inc.) and 1% penicillin/streptomycin (Wako Pure Chemical Industries, Ltd.).
The hyperthermia treatment was based on our previous study [11 (link)]. The MCF-7 cells were incubated at 42 °C with 5% CO₂ for 1 h using a standard incubator, and the MDA-MB-453 cells were incubated in the same way, but without CO₂.

Metformin (136-18662), glucose (049-31165), L-glutamine (074-00522), sodium pyruvate (199-03062), and sodium pyruvate solution (190-14881) were purchased from Fujifilm-Wako (Fujifilm-Wako Pure Medical Corporation, Miyazaki, Japan).

HepG2, Hep3B, and Huh-7 HCC cells were acquired from ATCC; JHH-4 and Huh-6 cells were acquired from the JRCB bank. Erlotinib, linsitinib, and lenvatinib were obtained from Selleck. Dulbecco’s Modified Eagle Medium (043-30085) and MEMα (135-15175) were purchased from Wako. Penicillin–streptomycin solution (× 100) (168-23191), 100 mmol/L sodium pyruvate solution (× 100) (190-14881), MEM non-essential amino acids solution (× 100)(139-15651), and 0.25 w/v% trypsin-1 mmol/L EDTA·4Na solution with phenol red (201-16945) were also purchased from Wako.