Vasilenko K P, Burova E B, Chupreta S V, Nikol'skył N N
Tsitologiia. 1997;39(2-3):150-8.
Dynamics of nuclear translocation of transcription factor Stat1 in human epidermoid carcinoma A431 cells in response to the epidermal growth factor (EGF) was examined by immunofluorescent microscopy and in cytoplasmic and nuclear extracts by Western blot. In has been shown that a prolonged presence of EGF induces a rapid tyrosine phosphorylation and nuclear translocation of Stat1 (within 5 min). The maximum amount of this protein in the nucleus was reached 30 min after the cell treatment to be maintained at the same level for 5 h. To study the dynamics of the export of Stat1 from the nucleus, a gentle treatment of cells with acetate buffer, pH 4.5, was used for extracting the surface-bound EGF. In this case, a complete dephosphorylation of Stat1 in the cytoplasm was observed in 30 min and the export of the Stat1 from the nucleus lasted for 1-6 h. These studies suggest the existence of an EGF-dependent dynamic equilibrium between the import and export of Stat1 in A431 cells.
通过免疫荧光显微镜以及蛋白质免疫印迹法检测人表皮样癌A431细胞中,转录因子Stat1在表皮生长因子(EGF)作用下的核转位动力学。研究表明,EGF的长期存在会诱导Stat1快速酪氨酸磷酸化和核转位(5分钟内)。细胞处理后30分钟,细胞核中该蛋白的量达到最大值,并在5小时内维持在同一水平。为了研究Stat1从细胞核输出的动力学,用pH 4.5的乙酸盐缓冲液温和处理细胞以提取表面结合的EGF。在这种情况下,30分钟内观察到细胞质中的Stat1完全去磷酸化,Stat1从细胞核的输出持续1 - 6小时。这些研究表明,A431细胞中Stat1的输入和输出之间存在EGF依赖的动态平衡。