Doxorubicin treatment causes a nuclear relocation of FOXO3a expression.
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The doxorubicin sensitive MCF-7 and resistant MCF-7-DoxR and MDA-MB-231 cells were treated with 1 µM doxorubicin. A) Cytoplasmic and nuclear protein lysates were prepared at the times indicated after doxorubicin treatment and protein expression levels were analyzed by Western blotting using antibodies against specific antibodies against FOXO3a, lamin B1 and and β-tubulin, B) The cells were cultured on sterile coverslips and treated for 16 h with or without 1 µM doxorubicin, before being fixed in 4% formaldehyde. FOXO3a expression was visualized with a specific rabbit polyclonal antibody against FOXO3a followed by the addition of ALEX488 (green) labeled anti-rabbit antisera. DAPI (blue) were also applied to visualize the nuclei.