Humec basal serum free media

4T1-Luc cells were a generous gift from Prof. G. Lazennec (Centre national de la recherche scientifique (CNRS), Montpellier, France). Cells were cultured in DMEM’s media (Sigma-Aldrich, Dorset, UK) supplemented with 10% FBS, 5 mL of MEM Non-Essential Amino Acids Solution (100×), 100 U/mL penicillin, 100 μg/mL streptomycin, 0.146 g/L l-glutamine (Sigma-Aldrich) and G418 (500 µg/mL).
Cells were split the day prior to injection and harvested at around 40–50% confluency.
MDA-MB-231 and MCF-7 were cultured in Dulbecco’s Modified Eagle Media (DMEM) without phenol red (Sigma-Aldrich) supplemented with 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, and 0.146 g/L l-glutamine (Sigma-Aldrich). Primary human mammary epithelial cells (pHMEC) were cultured in HuMEC basal serum free media (Thermofisher Scientific, Paisley, UK).

Human cancer cell line culture conditions were described previously (8 (link)). Briefly, cell lines were cultured with 5% CO₂ at 37°C. HEK293T, MCF-7, T47D, MDA-MB-361, ZR-75–1, MDA-MB-415, MDA-MB-134-VI, Hs578T and MDA-MB-231 were cultured in DMEM (Thermo Fisher) + 10% FCS. MDA-MB-436, MDA-MB-157, CAL51 and CAL120 cells were cultured in RPMI media with 10% FCS, 20 mM HEPES and 288 μl Insulin/100 ml. SUM159PT cells were cultured in Ham's F12 media with 5% FCS, 5 μg/ml insulin and 1 μg/ml hydrocortisone, HMLE cells in HuMEC basal serum free media (Thermo Fisher) and mesHMLE cells in Weinberg media (DMEM + F12 media with 5% FCS, 4 mg/ml insulin, 20 μg/ml EGF and 1 mg/ml hydrocortisone).
siRNAs were transfected with Lipofectamine RNAiMAX (Life Technologies) at 10 nM concentration following the manufacturer's protocol. DNA plasmid transfections were performed with Lipofectamine 2000 (Life Technologies) following the manufacturer's protocol.