Mccoy s medium

All cell lines were obtained from the American Type Culture Collection (Chicago, IL, USA), with the following exceptions: HCH-1 was from the laboratory of Dr. Linda Sandell at Washington University in St. Louis, MO, USA, and ASPS-1 was from the laboratory of Dr. Robert Shoemaker at the National Cancer Institute (Bethesda, MD, USA). SK-LMS-1, MNNG/HOS, MG-63, SK-UT-1 and SK-UT-1B cells were maintained in minimum essential media (Invitrogen, Waltham, MA, USA). Fuji, RD-ES, LUPI, NOS-1, HuO 9N2, HCH-1 and SK-ES cells were maintained in RPMI (Invitrogen). U-2 OS cells were maintained in McCoy's medium (Invitrogen). ASPS-1 cells were maintained in DMEM/F12 (Invitrogen). 293 T and SYO-1 cells were maintained in DMEM (Invitrogen). All media were supplemented with 10% fetal bovine serum (Invitrogen) and penicillin–streptomycin (Invitrogen). Chloroquine (used at 10 and 20 μM), 5-aza-dC (5 μM), necrostatin (10 μM) and ZVAD-FMK (100 μM) were obtained from Sigma Aldrich (St. Louis, MO, USA). Pepstatin A (used at 50 μM) and E64D (used at 25 μM) were obtained from Enzo Life Sciences (Farmingdale, NY, USA). ADI-PEG20 (used at 0.01, 0.05, 0.1, 0.5 and 1 μg/ml for dose-response curve, and at 1 μg/ml for all other in vitro experiments) was obtained from Polaris Pharmaceuticals, Inc. Lentiviral GFP-LC3 was a generous gift from Conrad Weihl (Washington University in St. Louis).^{27 (link)}

All the essential oils were provided by Bio-Jourdeness International Groups Co. Ltd. (Taichung Taiwan). Citronellol, geraniol, neryl acetate, and limonene were obtained from Tokyo Chemical Industry Co., Ltd. (Tokya, Japan). Purity of these compounds were above 99%, according to the GC and ¹H-NMR analysis. McCoy’s medium, fetal bovine serum (FBS), sodium pyruvate, penicillin, and streptomycin were obtained from Invitrogen (Carlsbad, CA, USA). 3-(4,5-Dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and dimethylsulfoxide (DMSO) were purchased from Sigma-Aldrich. Antibody against ACE2 was obtained from Arigo Biolaboratories (Hsinchu, Taiwan). Antibody against GAPDH was obtained from Cell Signaling Technology Inc. All other chemicals were reagent grade or HPLC grade and were supplied by either Merck or Sigma-Aldrich.

MDA-MB-231 breast carcinoma and SK-N-AS neuroblastoma cells were grown in DMEM medium (Invitrogen, Karlsruhe, Germany), T24 bladder carcinoma cells in McCoy's medium (Invitrogen) and Kym1 rhabdomyosarcoma cells in RPMI medium (Invitrogen) supplemented with 1% penicillin/streptomycin, 1% sodium pyruvate (both from Invitrogen) and 10% fetal calf serum (Invitrogen). The bivalent Smac mimetic BV6 that antagonizes XIAP, cIAP1 and cIAP2^{6 (link)} was kindly provided by Genentech Inc. (South San Francisco, CA, USA) and Enbrel by Pfizer (Berlin, Germany). Recombinant human TNFα was purchased from Biochrom (Berlin, Germany). All chemicals were obtained from Sigma (Deisenhofen, Germany) unless indicated otherwise.

Total bone marrow cells from the bilateral femurs of each mouse were eluted into McCoy's medium (Invitrogen, Carlsbad, CA, USA) using a 25-gauge needle. Spleens were mashed with the back of a 3 ml syringe plunger and filtered through a 70 μm cell strainers to collect the splenocytes.

MCF10A and SK-BR3 cells were obtained from ATCC (Manassas, VA). MCF10A-ERBB2 cells line is stably expressing wildtype ERBB2 (pBabe-Neu) as previously described [23 (link)]. MCF10A and MCF10A-ERBB2 cells were cultured in DMEM/F12 medium (Invitrogen, Carlsbad, CA) supplemented with 5% horse serum 20 ng/ml of EGF (Peprotech, Rocky Hill, NJ), 10 μg/ml of insulin (Sigma), 1 ng/ml of cholera toxin (Sigma), 100 μg/ml of hydrocortisone (Sigma), 50 units/ml of penicillin, and 50 μg/ml of streptomycin (Invitrogen); SK-BR3 in McCoy's medium (Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum. MTEC-Neu cells were kind gift from Tiffany Seagroves (University of Tennessee Health Sciences Center) and previously described [21 (link)]. LY294002 and U0126 were obtained from Calbiochem (San Diego, CA). Trametinib, lapatinib and dasatinib were purchased from Selleck Chemicals, LLC (Houston, TX).

Human cell lines were obtained from American Type Culture Collection (ATCC;CEM, Manassas, VA, USA) or Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ; Braunschweig, Germany) and cultured in DMEM medium or RPMI 1640 (Life Technologies, Inc., Eggenstein, Germany) or McCoy's medium (Invitrogen, Karlsruhe, Germany) supplemented with 10% fetal calf serum (FCS) (Biochrom, Berlin, Germany), 1 mM Sodium Pyruvate (Invitrogen, Karlsruhe, Germany) and 1% penicillin/streptomycin (Invitrogen, Karlsruhe, Germany). IFNα was obtained from Sigma-Aldrich (Taufkirchen, Germany), zVAD.fmk from Bachem (Heidelberg, Germany), Nec-1 from Merck (Darmstadt, Germany) and Enbrel from Pfizer (Berlin, Germany). The bivalent Smac mimetic BV6 was kindly provided by Genentech, Inc. (South San Francisco, CA, USA). All other chemicals were obtained from Sigma-Aldrich or Carl Roth (Karlsruhe, Germany) unless indicated otherwise.