Analysis of GFP-FOXO3a nuclear-cytoplasmic shuttling in ASTC-a-1 cells under growth factor stimulation

摘要

FOXO transcription factors are important regulators of cell survival in response to a variety of stimuli; among which are hypoxic stress; oxidative stress; and growth factor deprivation. Subcellular localization of FOXO proteins plays a major role in the regulation of their activity. In this study; using confocal imaging of the cells transfected with GFP-FOXO3a and fluorescence recovery after photobleaching technique; we visualized the dynamic nuclear translocation of GFP-FOXO3a in ASTC-a-1 cells under growth factor stimulus. In healthy cells; GFP-FOXO3a was well-distributed in the cytoplasm or widespread distributed in the cytoplasm and the nucleus but the cytoplasm was significantly more than the nucleus. Deprivation of growth factor; we monitored the nuclear localization of GFP-FOXO3a and the dynamic translocation of it from cytoplasm to nucleus. Interestingly; upon stimulation with growth factor in cells again; we visualized the dynamic nuclear exclusion of GFP-FOXO3a and cytoplasm distribution rapidly. In conclusion; these results demonstrated that FOXO3a can reversible shuttling between cytoplasm and nucleus upon stimulation with growth factor.