Breast Cancer Treatment Compound Evaluation

17β-estradiol (E₂), 4-OHT, anastrozole, insulin, hydrocortisone, cholera toxin, and DMSO were purchased from Sigma (St. Louis, MO). EGF was purchased from Peprotech (Rocky Hill, NJ), horse serum was from Invitrogen (Carlsbad, CA). TAM pellets (5 mg/pellet, 60-day release) and E₂ pellets (0.36 mg/pellet, 60-day release) were purchased from Innovative Research of America (Sarasota, FL, USA). Matrigel™ Matrix Growth Factor Reduced was purchased from BD Biosciences (Bedford, MA, USA). Anti-Raf-B, anti-p21, anti-Bcl-2, anti-Fibronectin and anti-β-actin antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). BreastDefend® (BD) was supplied by EcoNugenics, Inc. (Santa Rosa, CA, USA) and dissolved in DMSO at a concentration of 25 mg/ml then stored at −20 °C. The chemical composition of BD was previously published [26 (link)]. All other chemicals and reagents were of analytical grade.