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. Author manuscript; available in PMC: 2013 Dec 9.

Published in final edited form as: Mol Cancer Ther. 2011 Mar 10;10(5):10.1158/1535-7163.MCT-10-0836. doi: 10.1158/1535-7163.MCT-10-0836

A) Cells were treated for 24h with micromolar concentrations of ZOL (10, 15, 25 and 50μM). Then cell lysates were loaded in a 10% polyacrylamide gel (Criterion XT Bis-Tris precast Gel; Bio-Rad) and analyzed by immunoblot for MAPK, phospho-MAPK, Akt and phospho-Akt. Blots were reprobed with an antibody for β-actin as a control for protein loading and transfer. B) ZOL increases the localization of FOXO3a in the nuclear fraction of MDA-MB-231 cells. Cells were treated for 24h with increased concentrations of ZOL (10, 15, 25 and 50μM) and the nuclear and cytosolic extracts were prepared using the Nuclear Extract kit from Active Motif. Then total proteins (30μg) from both cell fractions were resolved by 10% acrylamide gel and analyzed by immunoblot for FOXO3a.

A) Cells were treated for 24h with micromolar concentrations of ZOL (10, 15, 25 and 50μM). Then cell lysates were loaded in a 10% polyacrylamide gel (Criterion XT Bis-Tris precast Gel; Bio-Rad) and analyzed by immunoblot for MAPK, phospho-MAPK, Akt and phospho-Akt. Blots were reprobed with an antibody for β-actin as a control for protein loading and transfer. B) ZOL increases the localization of FOXO3a in the nuclear fraction of MDA-MB-231 cells. Cells were treated for 24h with increased concentrations of ZOL (10, 15, 25 and 50μM) and the nuclear and cytosolic extracts were prepared using the Nuclear Extract kit from Active Motif. Then total proteins (30μg) from both cell fractions were resolved by 10% acrylamide gel and analyzed by immunoblot for FOXO3a.