HeLa cells were treated with 2 mM thymidine for 17 h, released for 7 h followed by treatment with 100 ng ml−1 nocodazole or 100 nM taxol for 15 h for inducing prometaphase arrest. MLN8054 (a gift from S. Gerber, Dartmouth, used at 1 μM to inhibit Aurora A and at 5 μM to inhibit both Aurora A and Aurora B), Aurora A inhibitor I (100nM, Selleckchem), ZM447439 (5μM, Tocris), hesperadin (100nM, Selleckchem), BI2536 (100nM, Selleckchem), flavopiridol (2 μM, Selleckchem), or roscovitine (50 μM, Millipore) were added for 45 mins. The cells were treated with MG132 (10 μM, Millipore) for 30 mins prior to adding the kinase inhibitors.
Aurora a inhibitor 1
Manufactured by Selleck Chemicals
Aurora A inhibitor I is a small molecule that inhibits the activity of the Aurora A kinase enzyme. Aurora A is a key regulator of cell division and is involved in the proper segregation of chromosomes during mitosis. This inhibitor functions by binding to and inhibiting the enzymatic activity of Aurora A, thereby disrupting the cell division process.
Automatically generated - may contain errors
Lab products found in correlation
Roscovitine, by Merck Group (1 mentions)
Flavopiridol, by Selleck Chemicals (1 mentions)
Hesperadin, by Selleck Chemicals (1 mentions)
Zm447439, by Bio-Techne (1 mentions)
Mg132, by Merck Group (1 mentions)
Bi2536, by Selleck Chemicals (1 mentions)
Egm mv single quots, by Lonza (1 mentions)
Nucleofector device with solution kit 5, by Lonza (1 mentions)
Tubacin, by Merck Group (1 mentions)
3 protocols using aurora a inhibitor 1
1
Prometaphase Arrest Induction Assay
2
Live Imaging of HUVECs on Fibronectin
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HUVECs were cultured and maintained in EBM medium supplemented with EGM-MV Single Quots (Lonza) at 37°C in 5% CO2. For live imaging, HUVECs were cultured at 50,000 cells/coverslip on 10 µg/ml fibronectin-coated glass coverslips (except for experiments using polyacrylamide substrates) and medium was supplemented with 25 µM Hepes, pH 7.2, and 30 U/ml Oxyrase. For Aurora A inhibition studies, cells were treated with 40 nM Aurora A Inhibitor I (Selleck Chemicals) for 60 min before imaging. Transfection of cDNAs and shRNA was performed using a Nucleofector Device with solution kit V (Lonza), setting A-034, and experiments were performed 6–10 h later to allow time for protein expression and/or shRNA knockdown.
3
Modulation of Signaling Pathways in Diverse Cell Types
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Shh small molecule agonist (SAG) was purchased from Sigma‐Aldrich (SML1314) and dissolved in sterile water. MEFs and hTERT‐RPE‐1 cells were treated with SAG (100 nM for MEFs and 200 nM and 500 nM for hTERT‐RPE‐1 cells) for 24 h after serum starvation with low serum medium (0.5% fetal bovine serum). Neurons were treated with 200 nM and 400 nM SAG for 48 h. Tubacin (SML0065, Sigma‐Aldrich) was dissolved in DMSO. MEFs, neurons, and hTERT‐RPE‐1 cells were treated with 1 μM Tubacin for 48 h, whereas fibroblasts were treated with 250 nM Tubacin for 24 h in low serum medium. Aurora A inhibitor I (TC‐S 7010, Selleckchem) was dissolved in DMSO, and a dose of 7 nM was used on neurons for 48 h.
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