Confluent MLO‐A5 cell cultures were treated with media supplemented with 0–10 μM N‐(benzyloxycarbonyl)‐Leu‐Leu‐Leucinal (MG132), lactacystin (Cayman Chemicals), Bortezomib (Velcade®) (Santa Cruz), and Withaferin‐A (Abcam, Cambridge UK) and then incubated at 37°C for 24 h. Confluent primary cell cultures were treated with media supplemented with ALLN, MG132, and lactacystin. Thereafter, protein/RNA was extracted from monolayers, as described below, for the determination of E11 protein/mRNA expression.
Withaferin a
Manufactured by Abcam
Sourced in United Kingdom
Withaferin‐A is a naturally occurring compound that has been studied for its potential biological activities. It is derived from the plant Withania somnifera. Withaferin‐A is commonly used in research applications to investigate its effects on various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Lactacystin, by Abcam (1 mentions)
Lactacystin, by Cayman Chemical (1 mentions)
Blebbistatin, by Selleck Chemicals (1 mentions)
Jasplakinolide, by Bio-Techne (1 mentions)
Nocodazole, by Abcam (1 mentions)
Y 27632, by Selleck Chemicals (1 mentions)
Celltiter glo assay, by Promega (1 mentions)
Paclitaxel, by Merck Group (1 mentions)
Nocodazole, by Merck Group (1 mentions)
Latrunculin a, by Santa Cruz Biotechnology (1 mentions)
Vinblastine, by Merck Group (1 mentions)
3 protocols using withaferin a
1
Proteasome Inhibitor Effects on E11 Expression
2
Tumor Cell Mechanics Modulation
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To soften tumor cells, HCC cells were pretreated with different doses of Y-27632 (2 and 20 μM; Selleckchem), blebbistatin (6 and 50 μM; Selleckchem), and cytochalasin D (0.1 and 1 μM; Selleckchem) for 48 h. To stiffen them, the cells were pretreated with 20 nM jasplakinolide (Tocris, #2792/100U) or 1 nM narciclasine (MedChemExpress) for 12 h. Nocodazole (1 and 6 μM; Abcam) and withaferin A (WFA; Abcam; 1, 2, and 5 μM) were used to disrupt microtubules and vimentin, respectively. The Wnt activator LiCl (2 μM) and inhibitor IWR-1-endo (0.2 μM) were used to activate and inhibit the Wnt/β-catenin pathway, respectively.
3
Cell Viability Assay of Anticancer Compounds
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VeroE6 cells were seeded into 96-well plates at a confluency of 7E+03 cells per well and treated for 6 h with serial 2-fold dilutions of the different compounds. For LatrunculinA, the highest used concentration was 5 μM, for all the other compounds a starting concentration of 50 μM was used. Nocodazole, paclitaxel and vinblastine were purchased from Sigma. LatrunculinA was obtained from Santa Cruz Biotechnology. WithaferinA was obtained from Abcam. DMSO was used as control. Cell viability was determined by quantitation of ATP concentration using the CellTiter-Glo assay (Promega) as recommended by the manufacturer. A Mithras LB 940 plate reader (Berthold Industries) was used to measure the luminescence. A compound concentration that reduced the ATP amounts more than 20% was considered cytotoxic.
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