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3 protocols using bibb 515

1

Evaluating Anti-Cancer Drug Synergies

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All chemicals were purchased from Sigma-Aldrich (St Louis, MO, USA) or Fisher Scientific (Pittsburgh, PA, USA) unless specified otherwise. Ficoll-Paque Plus was obtained from Amersham Biosciences (Piscataway, NJ, USA). The media for cell culture were purchased from GIBCO (Grand Island, NY, USA). The anti-CD-20 APC antibody was obtained from eBiosciences (San Diego, CA, USA). CellTiter 96® Non-Radioactive Cell Proliferation Assay kit (MTT) was purchased from Promega (Madison, WI, USA). Fludarabine was obtained from Teva Parenteral Medicines, Inc (Irvine, CA, USA). Rituximab was obtained from Genentech (South San Francisco, CA, USA). BIBB-515 and YM-53601 were purchased from Cayman Chemical Company (Ann Arbor, MI, USA). TAK-475 was obtained from Chemzon Scientific (Montreal, Quebec, Canada).
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2

Comprehensive Characterization of Small Molecules

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The identity and purity of key small molecules were authenticated by LC–MS ( Data S3). The following compounds were purchased from Cayman Chemicals: Ro 48–8071, BIBB 515, 20-hydroxycholesterol, 27-hydroxycholesterol, 22(S)-hydroxycholesterol, 22(R)-hydroxycholesterol, 24-hydroxycholesterol, 25-hydroxycholesterol, T0901317, GW 3965, WAY252623, and SR9238. Fenhexamid and CHIR-99021 were purchased from Sigma-Aldrich. VU 0255035 was purchased from Fisher. NB-598 was purchased from MedChemExpress. AGI-6780 and IWP-L6 were purchased from Selleck as DMSO stocks. Ciproxifan was purchased from Sigma Aldrich as a DMSO stock. The following standards were purchased from Toronto research chemicals: dioxidosqualene, mono-oxidosqualene, squalene. Details pertaining to all controls and GC-MS standards were reported previously.(Hubler et al., 2018 (link))
Methods and characterization of newly synthesized small molecules in included in Data S1-2.(Davis et al., 2003 (link), Wang et al., 1995 , Yang et al., 2018 (link), O’Keeffe et al., 2015 (link), Tomkinson et al., 1998 , Nakamura et al., 2009 , Maruoka et al., 1991 , Wu et al., 2010 , Emmons et al., 1989 , Shingate et al., 2013 , Shingate et al., 2011 , Jabbouri et al., 1991 , Nishitoba et al., 1988 , Jing et al., 2015 (link), Min et al., 1998 (link), Burns et al., 2018 (link))
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3

Comprehensive Cell Culture and Assay Protocols

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For cell culture, Dulbecco's modified Eagle's medium, Opti-MEM reduced serum medium, 0.05% trypsin–EDTA, and PBS were obtained from Gibco (Thermo Fisher Scientific); lipoprotein-deficient serum from fetal calf was purchased from Millipore Sigma. For transfection of DNA into HEK293T cells, transfection grade linear polyethyleneimine (PEI) HCl was purchased from Polysciences, Inc, and Fugene transfection agent was purchased from Promega. Phenylmethylsulfonyl fluoride and Roche complete EDTA-free protease inhibitor cocktail tablets were purchased from Thermo Fisher Scientific and Millipore Sigma, respectively. For coimmunoprecipitation, anti-FLAG M2 magnetic beads and Pierce anti-c-Myc magnetic beads were purchased from Millipore Sigma and Thermo Fisher Scientific, respectively. S-(5′-adenosyl)-l-methionine p-toluenesulfonate salt was purchased from Millipore Sigma, and CTP disodium salt hydrate was purchased from Acros Organics. For the SM and LS assays, NADPH tetrasodium salt, flavin adenosine dinucleotide disodium salt hydrate, 2,3-oxidoSqualene, lanosterol, and 2,6-di-tert-butyl-4-methylphenol (BHT) were purchased from Millipore sigma. Squalene was purchased from Thermo Fisher Scientific, and BIBB 515 was purchased from Cayman Chemical.
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