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Streptomycin

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Streptomycin is a broad-spectrum antibiotic derived from the bacterium Streptomyces griseus. It is commonly used in various lab applications to inhibit the growth of Gram-negative and Gram-positive bacteria.

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465 protocols using streptomycin

1

Culturing and Maintaining T. gondii Strains and Human Cell Lines

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T. gondii strains ME49 and Prugniaud were maintained in Vero cells in RPMI (Nacalai Tesque) supplemented with 2% heat-inactivated fetal bovine serum (FBS; JRH Bioscience), 100 U/mL penicillin, and 0.1 mg/mL streptomycin (Nacalai Tesque), as previously described (Ma et al., 2014 (link)). HFFs were maintained in RPMI (Nacalai Tesque) supplemented with 2% heat-inactivated FBS (JRH Bioscience), 100 U/mL penicillin, and 0.1 mg/mL streptomycin (Nacalai Tesque). IMR-32 cells were maintained in MEM (Nacalai Tesque) containing 10% heat-inactivated FBS, 1% non-essential amino acids (Nacalai Tesque), 100 U/mL penicillin, and 0.1 mg/mL streptomycin. A172 cells were maintained in DMEM (Nacalai Tesque) containing 10% heat-inactivated FBS, 100 U/mL penicillin, and 0.1 mg/mL streptomycin. U251-MG cells were maintained in EMEM (Nacalai Tesque) containing 10% heat-inactivated FBS, 100 U/mL penicillin, and 0.1 mg/mL streptomycin.
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2

Culturing Toxoplasma gondii and Cell Lines

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All T. gondii strains were maintained in Vero cells in RPMI (Nacalai Tesque) supplemented with 2% heat-inactivated FBS (JRH Bioscience), 100 U/ml penicillin, and 0.1 mg/ml streptomycin (Nacalai Tesque), as previously described (55 (link)). HAP1 cells were maintained in IMDM (Nacalai Tesque) containing 10% heat-inactivated FBS, and 100 U/ml penicillin, and 0.1 mg/ml streptomycin. HFFs, Huh7 cells were maintained in RPMI (Nacalai Tesque) containing 10% heat-inactivated FBS, and 100 U/ml penicillin, and 0.1 mg/ml streptomycin. HeLa cells and MEF cells were maintained in DMEM (Nacalai Tesque) containing 10% heat-inactivated FBS, 100 U/ml penicillin, and 0.1 mg/ml streptomycin.
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3

Cell Culture of Melanoma and Kidney Cell Lines

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The B16-F0 mouse melanoma cell line (cat. no. JCRB0202) was purchased from Japanese Collection of Research Bioresources (JCRB) Cell Bank and maintained in Eagle's Minimum Essential Medium (EMEM; FUJIFILM Wako Pure Chemical Corporation) supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich; Merck KGaA), 100 U/ml penicillin and 100 µg/ml streptomycin (both from Nacalai Tesque, Inc.) under mycoplasma-free conditions. Clone M3 (Cloudman S91) melanoma cell line was obtained from European Collection of Authenticated Cell Cultures (ECACC) and cultured in Ham's F10 medium (FUJIFILM Wako Pure Chemical Corporation) supplemented with 15% FBS, 2 mM glutamine (Nacalai Tesque, Inc.), 100 U/ml penicillin and 100 µg/ml streptomycin. 293T and 293FT cells were obtained from Invitrogen; Thermo Fisher Scientific, Inc. HEK-Blue™ TGF-β cells were purchased from InvivoGen. 293T, 293FT and HEK-Blue™ TGF-β cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Nacalai Tesque, Inc.) supplemented with 10% FBS, 100 U/ml penicillin and 100 µg/ml streptomycin. The cultured medium for 293FT cells was also supplemented with 1% non-essential amino acid solution (Nacalai Tesque, Inc.). All cell lines were cultured in a humidified incubator containing 5% CO2 at 37°C.
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4

Culture and Maintenance of Hepatic Cell Lines

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HepG2 cells and their derivatives (NTCP-expressing HepG2 cells [NTCP/HepG2]) were maintained in a Williams’ B culture medium (Gibco)-based PMM (primary hepatocyte maintaining medium), which contained 10% fetal bovine serum (FBS), 100 IU/mL penicillin, 100 µg/mL streptomycin and 0.25 µg/mL amphotericin B (Nacalai Tesque, San Diego, CA, USA), 50 µM hydrocortisone (Sigma-Aldrich), 5 µM dexamethasone, 5 µg/mL transferrin (Wako Pure Chemicals), 10 ng/mL EGF (ThermoFisher, Waltham, MA, USA), 5 µg/mL insulin (Sigma-Aldric), 5 ng/mL sodium selenite, 2 mM L-glutamine (Nacalai Tesque), and 0.5 mg/mL G418 (Nacalai Tesque). NTCP-expressing HepG2 cells were the kind gift of Dr. Watashi [43 (link),44 (link)] and were maintained in the same PMM but with the addition of 0.5 mg/mL G418 (Nacalai Tesque). HEK293T cells were cultured in Dulbecco’s modified Eagle media (DMEM) (high glucose) (Nacalai Tesque) supplemented with 10% FBS, 100 IU/mL penicillin, 100 µg/mL streptomycin, and 0.25 µg/mL amphotericin B (Nacalai Tesque). All the cells were maintained in a 5% CO2 incubator (Panasonic Health Care, Tokyo, Japan). Huh6 and HB611 cells were maintained in DMEM (low glucose) (Nacalai Tesque) with the same supplements as HEK293T. In the case of HB611, G418 (Nacalai Tesque) was added to the medium at 0.5 mg/mL.
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5

Preparation and Culture of Bone Marrow-Derived Macrophages

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Bone marrow-derived macrophages (BMMs) were prepared as described previously (Ichinohe et al., 2009 (link)). In brief, bone marrow was obtained from the tibia and femur by flushing with Dulbecco’s modified Eagle’s medium (DMEM; Nacalai Tesque). Bone marrow cells were cultured for 5 days in DMEM supplemented with 30% L929 cell supernatant containing macrophage colony-stimulating factor, 10% heat-inactivated fetal bovine serum (FBS), and L-glutamine (2 mM) at 37°C/5% CO2. HEK293FT cells (a human embryonic kidney cell line) and HeLa cells (a human epithelial carcinoma cell line) were maintained in DMEM supplemented with 10% FBS, penicillin (100 units/ml), and streptomycin (100 μg/ml) (Nacalai Tesque). MDCK cells (Madin-Darby canine kidney cells) and HT-1080 cells (a human fibrosarcoma cell line) were grown in Eagle’s minimal essential medium (E-MEM; Nacalai Tesque) supplemented with 10% FBS, penicillin (100 units/ml), and streptomycin (100 μg/ml) (Nacalai Tesque).
Influenza A virus strain A/PR8 (H1N1) was grown at 35°C for 2 days in the allantoic cavities of 10-day-old fertile chicken eggs (Ichinohe et al., 2009 (link)). The viral titer was quantified in a standard plaque assay using MDCK cells (Pang et al., 2013 (link)).
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6

Culturing T. gondii and Cell Lines

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C57BL/6NCrSlc (C57BL/6N) mice were purchased from Japan SLC, Inc. All animal experiments were conducted with the approval of the Animal Research Committee of Research Institute for Microbial Diseases in Osaka University. Pru strains of T. gondii were maintained in Vero cells by biweekly passage in RPMI 1640 (Nacalai Tesque) supplemented with 2% heat-inactivated FBS (Gibco), 100 U/ml penicillin, and 0.1 mg/ml streptomycin (Nacalai Tesque). B16F10, Plate-E, and 293T cells were maintained in DMEM (Nacalai Tesque) supplemented with 10% heat-inactivated FBS, 100 U/ml penicillin, and 0.1 mg/ml streptomycin (Nacalai Tesque).
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7

Mouse model for Toxoplasma gondii study

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C57BL/6NCrSlc (C57BL/6N) mice were purchased from SLC. Mice lacking IFN-γR have been previously described (30 (link)). All animal experiments were conducted with the approval of the Animal Research Committee of Research Institute for Microbial Diseases at Osaka University. RHΔhxgprtΔku80 and its derivatives of T. gondii were maintained in Vero cells by bi-weekly passage in RPMI (Nacalai Tesque) supplemented with 2% heat-inactivated fetal calf serum (FCS; JRH Bioscience), 100 U/mL penicillin and 0.1 mg/mL streptomycin (Nacalai Tesque). Mouse embryonic fibroblasts were maintained in Dulbecco’s modified Eagle medium (DMEM; Nacalai Tesque) supplemented with 10% heat-inactivated FCS, 100 U/mL penicillin, and 0.1 mg/mL streptomycin (Nacalai Tesque).
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8

Cell Line Cultivation and Maintenance

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TC71, SK‐ES‐1, RD‐ES, RD, and human epithelial kidney 293T cell lines were provided by the laboratory of Professor Tohru Sugimoto (University of Miyazaki, Kyoto Prefectural University of Medicine). The SU‐CCS‐1 cell line was kindly supplied by Dr Alan L. Epstein (Keck School of Medicine, University of Southern California), and the HS‐Os‐1 cell line was obtained from Riken Bioresource Center (Tsukuba, Japan). Human fibroblasts were obtained through the culture of mononuclear cells isolated from the bone marrow of healthy volunteer with informed consent. Except for HS‐Os‐1 and 293T, all cells were grown in RPMI (Sigma‐Aldrich) with 10% fetal bovine serum (Sigma‐Aldrich), 100 U/mL penicillin, and 100 µg/mL streptomycin (Nacalai Tesque Inc). HS‐Os‐1 and 293T cells were maintained in Dulbecco's Modified Eagle's Medium (Sigma‐Aldrich) with 10% fetal bovine serum (Sigma‐Aldrich), 100 U/mL penicillin and 100 µg/mL streptomycin (Nacalai Tesque Inc). SK‐ES‐1 and RD‐ES were cultured on collagen‐coated dishes or flasks.
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9

Diverse Cell Lines for Viral Propagation

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The human cell lines A549, ACHN, ADR-RES, H23, OVCAR-3, OVCAR-4, OVCAR-5, SK-OV3, SK-MEL28, and T-47D were obtained from Dr. Toru Okamoto (the Walter & Eliza Hall Institute). Mouse fibroblast L929, mouse embryonic fibroblast (MEF), baby hamster kidney BHK, African green monkey kidney cell lines (Vero, CV-1, and MA104), canine kidney MDCK, and quail fibrosarcoma QT-6 cell lines were obtained from the American Type Culture Collection. Bovine kidney epithelial MDBK cell line and swine kidney SK-6 cell line [69 (link)] were kindly provided from Dr. Tomokazu Tamura. These human and animal cell lines were grown in Dulbecco’s modified Eagle’s medium (DMEM; Nacalai Tesque) supplemented with 5% fetal bovine serum (FBS; Gibco), 100 units/mL penicillin, and 100 μg/mL streptomycin (Nacalai Tesque). BHK-T7 cells were cultured in DMEM supplemented with 5% FBS, 100 units/mL penicillin, 100 μg/mL streptomycin, and 8 μg/mL puromycin (Sigma-Aldrich) [70 (link)]. DemKT1 and YubFKT1 cell lines were kindly provided from Dr. Ken Maeda [49 (link)] and grown in Roswell Park Memorial Institute 1640 medium (RPMI; Nacalai Tesque) supplemented with 5% FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin. Wild-type rsMB was generated previously using a reverse genetics system [45 (link)]. Viruses were propagated in L929 cells. Viral titers were determined by plaque assay using L929 monolayers.
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10

Establishing Canine EGFR and HER2 Overexpressed CHO Cells

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CHO-K1 and a canine osteosarcoma cell line D-17 were purchased from the American Type Culture Collection (Manassas, VA, USA). Stable dog EGFR-overexpressed CHO-K1 (CHO/dEGFR) [26 (link)] and dog HER2-overexpressed CHO-K1 (CHO/dHER2) [27 (link)] were established as described previously. CHO-K1, CHO/dEGFR, and CHO/dHER2 were cultured in RPMI-1640 medium (Nacalai Tesque, Inc., Kyoto, Japan), containing 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Inc., Waltham, MA, USA), 100 μg/mL streptomycin, 100 units/mL of penicillin, and 0.25 μg/mL amphotericin B (Nacalai Tesque, Inc.). D-17 was cultured in MEM medium (Nacalai Tesque, Inc.), supplemented with 10% FBS, 1 mM of sodium pyruvate, 100 units/mL of penicillin, 100 μg/mL streptomycin, and 0.25 μg/mL amphotericin B.
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