Culturing and Characterizing TNBC and CRPC Cell Lines

Human BT-549 TNBC cells (ATCC) were cultured in RPMI1640 medium (Thermo Fisher Scientific, Waltham, MA, USA) containing 10% fetal bovine serum (FBS; GEMINI Bio-Products, West Sacramento, CA, USA), 100 μg/ml streptomycin, 100 U/ml penicillin and 10 μg/ml insulin. Human MDA-MB-468 TNBC cells (ATCC) were cultured in Leibovitz’s L-15 medium (Thermo Fisher Scientific) containing 10% FBS. Human SUM149 BRCA1 mutant TNBC cells (ATCC) were grown in Ham’s F-12 medium (Corning, Manassas, VA, USA) supplemented with 10 mM HEPES, 5% FBS, 100 μg/ml streptomycin, 100 U/ml penicillin, 5 μg/ml insulin and 1 μg/ml hydrocortisone. Human LNCaP-AI CRPC cells^{29 (link)} were cultured in phenol red-free RPMI1640 medium (Thermo Fisher Scientific) containing 10% charcoal-stripped FBS (Millipore Sigma, Burlington, MA, USA). DU-145 CRPC cells (ATCC) were cultured in RPMI1640 medium (Corning Life Sciences, Corning, NY, USA) containing 10% heat-inactivated FBS. Cells were treated with the NF-κB inhibitor BAY11-7082 (S2913; Selleckchem, Houston, TX, USA) and MUC1-C inhibitor GO-203^{12 (link)}. Cells were maintained in culture for 3–4 months. Authentication of the cells was performed by short tandem repeat (STR) analysis. Cells were monitored for mycoplasma contamination using the MycoAlert Mycoplasma Detection Kit (Lonza, Rockland, ME, USA).