The human lung cancer cell lines A549, H358, and H460 and the lung epithelial cell line 16HBE were purchased from the Cell Bank at the Shanghai Institutes for Biological Sciences of the Chinese Academy of Sciences and were cultured in 1640 medium supplemented with 10% foetal bovine serum and 1% double antibiotics, as instructed by their provider. HEK293T and HeLa cells were cultured in DMEM and 10% foetal bovine serum (ExCell Bio, Shanghai, China) supplemented with 1% double antibiotics. All cells were maintained at 37°C with 5% CO2. The cells were treated with Importazole (ab146155) [27 (link)], Leptomycin B (ethanol solution, ab120501) or Bufexamac (S3023, Selleck, Shanghai, China) for the indicated times.
Importazole
Manufactured by Selleck Chemicals
Sourced in United States
Importazole is a laboratory equipment designed for the isolation and purification of biomolecules. It utilizes a proprietary chromatographic technique to separate and concentrate target analytes from complex mixtures. The core function of Importazole is to facilitate the extraction and purification of substances of interest, supporting various research and analytical applications.
Automatically generated - may contain errors
Lab products found in correlation
Rnase free dnase, by Thermo Fisher Scientific (2 mentions)
Facscalibur, by BD (2 mentions)
Leptomycin b, by Selleck Chemicals (1 mentions)
Foetal bovine serum (fbs), by ExCell Bio (1 mentions)
Dulbecco modified eagle medium (dmem), by ExCell Bio (1 mentions)
Cycloheximide (chx), by Thermo Fisher Scientific (1 mentions)
Cytochalasin b, by Bio-Techne (1 mentions)
2 deoxyglucose, by Merck Group (1 mentions)
Sodium azide, by Merck Group (1 mentions)
Leptomycin b, by Merck Group (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Nocodazole, by Merck Group (1 mentions)
4 protocols using importazole
1
Cell Lines and Treatment Conditions
2
Modulation of Cellular Trafficking Pathways
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Cells were treated with cytochalasin B (Tocris Biosciences, 20 μM, 10 min) and latrunculin B (MilliporeSigma, 1 μM, 10 min) to depolymerize actin, and nocodazole (MilliporeSigma, 1 μg/mL, 15 min) to depolymerize microtubules. Cells were treated with leptomycin B (Sigma-Aldrich, 20 ng/mL, 48 hr) to inhibit CRM1-mediated export. Importazole (Selleck Chemicals, 40 μM, 1 hr) was used to inhibit Importin β activity. Cells were treated with guanosine (TCI America, 100 μM, 48 hr) to supplement GTP synthesis. Cells were treated with cycloheximide (ThermoScientific, 350 μM, 2 hr) to inhibit protein translation. Cells were treated with MPA (ThermoScientific, MPA, 1.8 μM, 48 hr) to deplete GTP. Cells were treated with sodium azide (Sigma-Aldrich, 10mM, 20 min) and 2-deoxyglucose (Sigma-Aldrich, 6mM, 20 min) to deplete ATP. Control cells were treated with equal volumes of media. Nuclear import of GR-GFP was induced by the addition of dexamethasone (Sigma-Aldrich, 1 μM, 15 min).
3
Cell Cycle Analysis by Flow Cytometry
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PC3 or C42B cells were seeded in a 6-wells plate (0.8 million cells per well) and transfected with siRNA (Invitrogen) for 48 hours or treated with Importazole (Selleckchem, Houston, TX, USA) for 24 hours. The post-treatment cells were collected and fixed with 70% ethanol. After 2 washes with PBS, the cells were stained with propidium iodide (PI) solution (20 μg/ml PI (Roche, Basel, Switzerland) in PBS containing 0.1% Triton-100 (Sigma-Aldrich-Aldrich) and 0.2 mg/ml DNAse-free RNAse (Thermo fisher scientific)) and incubated at 37℃ for 15 minutes. The stained cells were washed with PBS for once and re-suspended in 500 μl PBS. Flow cytometry was performed on a BD FACSCalibur flow cytometer (Becton Dickinson, Franklin Lakes, NJ, USA). The results were interpreted and displayed using Flowjo software (Flowjo, Ashland, OR, USA).
4
Cell Cycle Analysis by Flow Cytometry
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