Mda mb 231

The normal mammary epithelial cell line MCF-10 A and the breast cancer cell lines MDA-MB-231, MCF-7, SK-BR3, T47D and Cal51 were obtained from ATCC (American Type Culture Collection, Manassas, USA). MCF-10 A was cultured in MCF-10 A cell specific medium (Procell, China). MCF-7, T47D, Cal51 and MDA-MB-231 were cultured in DMEM (Gibco, Grand Island, USA). All medium included 10% fetal bovine serum (FBS, NEWZERUM, Australia) and 1% penicillin/streptomycin (Gibco, Grand Island, USA). All media contained 10% fetal bovine serum (FBS, NEWZERUM, Australia) and 1% penicillin/streptomycin (Gibco, Grand Island, USA). Cells were cultured at 37 °C in a humidified cell incubator with a 5% CO2 atmosphere. The SMYD4 plasmids and corresponding vectors as well as the SMYD4 siRNA kits were purchased from RiboBio (Guangzhou, China). MYH9 plasmids and corresponding vectors were purchased from GeneCopoeia (Guangzhou, China). The siRNA kits of MYH9 were obtained from Keyybio (Shandong, China). Transfection was performed using Lipofectamine 3000 (Invitrogen, California, USA) according to the manufacturer’s recommendations. Lentiviruses (RiboBio, China) were used to infect CAL-51 cells and generate stable SMYD4-overexpressed cells according to the manufacturer’s protocol. The siRNAs oligonucleotides are listed in Supplemetary information: Table S1.

The MDA-MB-231 and Hs578T human breast cancer cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA). MDA-MB-231 LM1 cell was provided by SJ Lee (Fibrosis and Cancer Targeting Biotechnology, Seoul, Korea). All cell lines were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. Cells were incubated at 37 ℃ in a humidified atmosphere with 5% CO₂.

MDA-MB-231 and 4T1 cell lines were from the American Type Culture Collection (ATCC). SUM149 cell line was kindly gifted from Dr. Debeb (MD Anderson Cancer Center) and SUM159 cell line was purchased from BioIVT and also provided from Asterand Bioscience by Dr. Dave. c-Myc overexpressing MTB-TOM cell was provided by Dr. Goga (UCSF). Rho0 cell and TNBC cybrid cells, C-A1N4 and C-SUM159, were maintained as previously published.⁶ (link) MDA-MB-231, 4T1, SUM149, SUM159, and TNBC cybrid cells were cultured in DMEM medium containing 5% FBS and 1% penicillin-streptomycin and Rho0 cell was cultured in DMEM medium containing 5% FBS, 1% penicillin-streptomycin and 50 ng/ul uridine. MTB-TOM cell was maintained in DMEM medium containing 5% tet-free FBS and 1% penicillin-streptomycin. All cells were incubated at 37^oC with 5% CO₂.

The human breast cancer cell lines MDA-MB-231 (ATCC) were cultured with RPMI1640 containing 10% FBS, 2 mm L-glutamine, and 1% Penicillin (10000 U/mL)-Streptomycin (10000 μg/mL) at 37 °C with 5% CO₂. MCF-10A cells (ATCC) were incubated with DMEM/F12 medium including 10% FBS, 2 mm L-glutamine, 10 μg/mL human insulin and 1% Penicillin (10000 U/mL)-Streptomycin (10000 μg/mL) at 37 °C with 5% CO₂. Quercetin (CAS No: 117–39-5, ≥ 98% purity), Luteolin (CAS No: 491–70-3, ≥ 98% purity), Baicalein (CAS No: 491–67-8, ≥ 98% purity) were purchased from Sigma Aldrich and then dissolved in 0.5% dimethyl sulfoxide (DMSO) for drug administration. The composite solution of the three compounds (QLB solution) was prepared by mixing Quercetin, Luteolin, and Baicalein at ratio of 1.5:1.5:1, to match their respective levels in the TSGJ solution.