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6 protocols using tigecycline

1

Tigecycline Susceptibility Determination

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Susceptibility to Tigecycline was determined using the micro‐broth dilution method. Tigecycline was purchased from Selleck Chemicals (Houston, TX; Lot: S140303) and was used at a concentration ranging from 0·06 to 128 μg ml−1. Cation‐adjusted Mueller–Hinton broth was purchased from Bio‐Kont Co., Ltd (Wenzhou, China; Lot: HB6231‐1). The ratio of the strain to the cation‐adjusted Mueller–Hinton broth was 1 : 200, with 100 μl added to each well of a drug sensitivity plate. The plate was placed in an incubator with 5% carbon dioxide at 35°C overnight, and the results were observed on the second day.
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2

Anticancer Drug Screening Compounds

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Tigecycline (S1403), Cobimetinib (S8041), Gefitinib (S1025) and 2-Deoxy-d-glucose (S4701) were purchased from Selleck Chemicals. Trametinib (HY-10999) was purchased from MedChemExpress.
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3

Evaluating Adriamycin and Tigecycline Synergy

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Adriamycin (99.82% purity, No. S1208) and Tigecycline (99.86% purity, No. S1403) were purchased from Selleckchem (Houston, TX, USA), while DMSO (99.9% purity) was purchased from Solarbio (Beijing, China).
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4

Comparison of Tetracycline Derivatives

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Doxycycline hyclate, minocycline HCl and tigecycline were obtained from Selleck Chemicals, Houston, TX, USA. EtOH (200 proof, Decon Labs, King of Prussia, PA, USA) was used in all experiments. For some studies, saccharin sodium salt hydrate (Sigma-Aldrich, St. Louis, MO, USA) was used. Vehicle for all drugs was physiological saline and drugs were given by intraperitoneal injection (ip) in a volume of 10 ​ml/kg body weight.
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5

Dose-Dependent Drug Screening Assay

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For drug dose response assays, 5000 cells were plated in each well of a 96-well plates. Drugs were serially diluted in base media and applied to a total volume of 100µL per well. After four days of culture, viability was assessed by the addition of 30 µL/well of Celltiter-Glo (Promega, cat#G7572) and measuring luminescence on a Synergy H1 microplate reader (Biotek). Drugs were purchased as follows: irinotecan (Selleckchem, cat#S221); AZD1775 (Selleckchem, cat#S1525); ribociclib (Selleckchem, cat#S7440); carboplatin (Sigma, cat#C2538); etoposide, (Selleckchem, cat#S1225); alisertib (Selleckchem, cat#S1133); ABT-199 (Selleckchem, cat#S8048); GSK128 (Selleckchem, cat#S7061); tigecycline (Selleckchem, cat#S1403); talazoparib (Selleckchem, cat#S7048); VX-970 (Selleckchem, cat#S7102).
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6

Ovarian Cancer Cell Line Characterization

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OVCAR-8 and OVCAR-5 cells were kind gifts from D. Scudierio (NCI, National Institutes of Health). The PEA1 cell line was from Sigma-Aldrich. The BRCA1-mutant COV362 cell line was a gift from Robert van Waardenburg, University of Alabama Birmingham. The cells were cultured in RPMI-1640 medium (Corning) supplemented with 8% fetal bovine serum (Millipore), and maintained in a humidified 37°C, 5% CO2 incubator. All cells were authenticated by autosomal STR profiling (University of Arizona Genetics Core). All cell lines were free of Mycoplasma contamination as determined testing with a MycoAlert Mycoplasma Detection Kit (Lonza). VLX600 was obtained from Cayman Chemical. Tigecycline and WZB117 were obtained from Selleck Chemicals.
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