For the MMTV-PyMT model, animals were maintained at the Dana-Farber Cancer Institute (Protocol 10-067). For primary screens, tumors were collected from different mammary fat pads and pooled to get appropriate numbers of cells. For animal treatments, MMTV-PyMT cells that were extracted from mice and not cultured in vivo were injected along with Matrigel (Corning) into the mammary fat pad of syngenic FVB/NJ mice as described previously (40). Seven to 15 mice were used for each in vivo treatment arm. Dasatinib (ApexBio) and 17-DMAG (ApexBio) were intraperitoneally dosed at 10 mg/kg. AZD2014 (ApexBio) was dosed via oral gavage once daily at 15 mg/kg. Lapatinib (SelleckChem) was dosed via oral gavage at 50 mg/kg. Sunitinib (ApexBio) was dosed via oral gavage at 50 mg/kg. Navitoclax (MedChemExpress) was dosed via oral gavage at 100 mg/kg. All treatments were performed daily, 5 days a week for 2 weeks.

With regard to breast PDX tumors, DF-BM355 tumors were previously treated with compounds in vivo (32) and were expanded as previously described (32). For colon PDX tumors, tissues to establish PDX models were obtained according to Institutional Review Board (IRB)–approved research protocols (14-030). Fresh primary colorectal cancer biopsies were first incubated in an antibiotic cocktail of penicillin/streptomycin/amphotericin B/ciprofloxacin for 1 to 2 hours and implanted into the flanks of 5-weeks-old female nude mice (Nu/Nu; Taconic). When the xenografts reached ~200 mm, three mice were sacrificed, and the tumors were harvested and serially passaged as subcutaneous implants of tumor fragments about 2 to 3 mm in diameter.

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