The human hepatoma-derived cell line HuH-7 was obtained from the Japanese Cancer Research Resources Bank (Osaka, Japan). OR6 cells derived from HuH-7 cells with the stable transfection of the full-length genotype 1 replicon containing the Renilla luciferase gene were selected by neomycin, ORN/C-5B/KE (23 (link)), were used to examine the anti-HCV effect of IFN-α. The cells were cultured and maintained in Dulbecco's modified Eagle's medium (DMEM; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) containing 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Inc., Waltham, MA, USA) and 1% antibiotics (ampicillin/streptomycin) in 5% CO2 at 37°C. ECM (type I collagen, laminin, type IV collagen or fibronectin)-coated dishes (Cosmo Bio, Tokyo, Japan) were used for cell culture to investigate the differences in cell signaling between cells cultured on ECM-coated dishes and those cultured on non-ECM-coated dishes, which had hydroxyl and carboxyl groups on the surface to facilitate cell adhesion (cat. no. 150687; Thermo Fisher Scientific, Inc.).
Ampicillin streptomycin
Manufactured by Thermo Fisher Scientific
Sourced in United States
Ampicillin/streptomycin is an antibiotic solution used in cell culture media to prevent bacterial contamination. It contains the antibiotics ampicillin and streptomycin.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Corning (1 mentions)
T75 flask, by Corning (1 mentions)
Hsdhan aura strain, by Inotiv (1 mentions)
Vero e6 cell, by American Type Culture Collection (1 mentions)
C2c12 cell, by American Type Culture Collection (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Cytiva (1 mentions)
Emt antibody sampler kit, by Cell Signaling Technology (1 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
P erk tyr202 204, by Cell Signaling Technology (1 mentions)
Ap24534, by Selleck Chemicals (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Hucct1, by National Collection of Authenticated Cell Cultures (1 mentions)
Hccc9810, by National Collection of Authenticated Cell Cultures (1 mentions)
Croton oil, by Merck Group (1 mentions)
Dulbecco s modified eagle medium nutrient mixture f 12 dmem f 12, by Thermo Fisher Scientific (1 mentions)
10 protocols using ampicillin streptomycin
1
HuH-7 Cells for Anti-HCV Screening
2
Cytotoxicity Evaluation of Synthesized LDHs
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The cytotoxicity of the synthesized LDHs (56 and 88 nm) was evaluated using trypan blue staining and the resazurin assay. HEK-293T cells were grown in DMEM (Corning Inc, Corning, NY, USA) supplemented with ampicillin/streptomycin (Thermo Fisher Scientific, Waltham, MA, USA) and 10% heat-inactivated fetal bovine serum (Gibco®, Life Technologies Corporation, Grand Island, NY, USA) (complete DMEM) at 37 °C and 5% CO2 using T75 flasks (Corning) until reaching confluence. One day before the cytotoxicity evaluation, 5 × 104 cells were seeded in triplicate in a 24-well culture plate. As control experiments, each plate had cells treated with the vehicle alone (PBS) or H2O2 (40 mM). The cells were subsequently exposed to different concentrations of LDH (from 6.5 to 500 μg/mL) for 48 h and 72 h under the aforesaid culture conditions. Afterward, the resazurin-based cell viability was estimated. For this purpose, the cells treated for 48 h and 72 h with the respective LDH concentrations were exposed to resazurin at 30 μg/mL for 3 h, and the fluorescence (560 nm/590 nm) was recorded in a FlexStation II scanning fluorimeter (Molecular Devices LLC, San Jose, CA, USA).
3
Isolation and Characterization of Mouse Lung Vascular Endothelial Cells
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MLVECs were isolated as previously described (26 (link)). Briefly, peripheral and subpleural tissue was cut into small pieces (1-mm3) and tissue fragments were placed into 60-mm cell culture dishes. DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 20% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.) was added as culture medium. Following 48 h incubation at 37˚C, the tissue was removed and MLVECs were cultured in DMEM supplemented with 10% FBS (Gibco) and 0.01% ampicillin/streptomycin (Gibco; Thermo Fisher Scientific, Inc.) at 37˚C with 5% CO2. MLVECs were characterized by their cobblestone morphology and stained with factor VIII-related antigen (cat. no. sc-53466, 1:100, Santa Cruz Biotechnology, Inc.) at 4˚C for one night as previously described (27 (link)). Cells after 3-4 passages were used in subsequent experiments.
4
Hamster Vero E6 Cell Culture
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The Syrian golden hamsters (HsdHan:AURA strain) were purchased from Envigo (Indianapolis, IN). Vero E6 cells, an African green monkey kidney epithelial cell line (ATCC, Manassas, VA, USA), were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco/Thermo Fisher, Waltham, MA, USA) with 10% fetal bovine serum (FBS; HyClone Laboratories, South Logan, UT) plus 1% ampicillin/streptomycin (Gibco). The authenticity of Vero E6 cells was verified using Short Tandem Repeat profiling by ATCC. The cells were tested negative for mycoplasma.
5
Differentiation and LCA Treatment of C2C12 Myoblasts
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C2C12 cells (ATCC, Manassas, USA) were cultured in growth medium (DMEM; Gibco, Carlsbad, USA) supplemented with 10% fetal bovine serum (FBS; Gibco) and 100 IU/mL ampicillin-streptomycin (Gibco) in a constant-humidity atmosphere (5% CO
2 and 37°C). For differentiation into myotubes, C2C12 cells were plated in 12-well plates at a density of 5×10
4 cells/well. When C2C12 cells reached 80% confluence, the medium was changed to differentiation medium supplemented with 2% horse serum (HyClone, Logan, USA) and cultured for another 3 days. Then, C2C12 cells were treated with 50 μM LCA (MCE) for 3 days. After treatment, C2C12 cells were harvested for western blot analysis and qRT-PCR.
2 and 37°C). For differentiation into myotubes, C2C12 cells were plated in 12-well plates at a density of 5×10
4 cells/well. When C2C12 cells reached 80% confluence, the medium was changed to differentiation medium supplemented with 2% horse serum (HyClone, Logan, USA) and cultured for another 3 days. Then, C2C12 cells were treated with 50 μM LCA (MCE) for 3 days. After treatment, C2C12 cells were harvested for western blot analysis and qRT-PCR.
6
Cell Line Maintenance and Treatment
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Nthy‐ori 3‐1 (abbreviated as Nthy thereafter), TPC‐1 and BCPAP cells were purchased from the Chinese Academy of Science Cell Bank. For cell line authentication, short tandem repeat profiling was performed before the beginning of experiment. All cell lines were maintained in RPMI‐1640 (Gibco) media supplemented with10%FBS (Gibco) and 1% ampicillin/streptomycin (Gibco) and cultured at 5% CO2 and 37°C. All cell lines were passaged <10 times. Cells were treated with 2‐DG (2.5 mM), MG132 (10 μM), CHX (20 μg/ml) and oligomycin (0.1 mM) at indicated times, respectively.
7
Vero E6 Cell Culture Protocol
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Vero E6 cells, an African green monkey kidney epithelial cell line (ATCC, Manassas, VA, USA), were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco/Thermo Fisher, Waltham, MA, USA) with 10% fetal bovine serum (FBS; HyClone Laboratories, South Logan, UT) plus 1% ampicillin/streptomycin (Gibco). The authenticity of Vero E6 cells was verified through STR profiling by ATCC. The cells were tested negative for mycoplasma.
8
FGF1 Signaling Pathway in ICC Cell Lines
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The ICC cell lines RBE, HuCCT1 and HCCC9810 were all purchased from Cell Bank of the Chinese Academy of Sciences (Shanghai, China) and cultured in the RPMI‐1640 medium supplemented with 10% foetal bovine serum (Gibco, Waltham, MA) and 1% ampicillin/streptomycin (Gibco) in 5% CO2 resuscitation. Human recombinant FGF1 was purchased from PeproTech Company, and the inhibitor AP24534 was purchased from Selleck, the primary antibodies of SPRY1‐4, pan‐phospho‐Tyr, and β‐actin were bought from Santa Cruz Biotechnology (Santa Cruz, CA), antibodies of FGFR2, Grb2, pFRS2‐Tyr436, p‐ERK‐Tyr202/204 and the EMT antibody sampler kit were purchased from the Cell Signaling Technology (Danvers, MA), antibody of p‐FGFR2‐Tyr769 was from Biorbyt Company (Cambridge, UK).
9
Inflammatory Response Assay Protocol
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Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12 (DMEM-F12), ampicillin/streptomycin, and phosphate-buffered saline (pH 7.4) were purchased from Gibco Invitrogen (Carlsbad, CA). Trypsin-EDTA (0.25%) and fetal bovine serum were obtained from HyClone Thermo Scientific (Logan, UT). Triton X-100 was acquired from Research Organics (Cleveland, OH). Lipopolysaccharides (LPS) from Salmonella enterica serotype typhimurium L7261, as well as croton oil, indomethacin, isorhamnetin standard, and formic acid solution HPLC grade were purchased from Sigma-Aldrich (St. Louis, MO). Chromatography grade water and methanol (VWR International LLC, West Chester, PA) were used for high-pressure liquid chromatograph equipped with a photodiode array detector (HPLC-PDA) and liquid chromatograph/mass selective detector time-of-flight (LC/MSD TOF) analysis.
10
Culturing Human Liver Cancer Cells
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Human HCC cells PLC/PRF/5 and SK-Hep-1 (ATCC), and HuH-7 (Japan Health Sciences Foundation) were cultured in 44% DMEM, 44% Ham's F-12, 10% FCS, 1% Glutamine, and 1% Ampicillin/streptomycin (Invitrogen).
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