CIL:13886

Constitutive targeting of Tem1 to the spindle pole body (SPB) in metaphase cells (CIL# 13883) and anaphase cells (this image). tem1Δ::GAL-UPL-TEM1 cells expressing eGFP-CNM67–TEM1 from a CEN plasmid were grown on 2% raffinose/2% galactose. Image shows localization of Cnm67-Tem1 (eGFP, green) in anaphase after cells were transferred to medium with 2% glucose. Tem1 normally localizes preferentially to the SPB that enters the daughter cell during anaphase (CIL# 13882, 13885). Cnm67 is an integral component of the outer plaque of the SPB and the Cnm67-Tem1 fusion is found to be symmetric from SPB duplication until the end of mitosis. Nuclear morphology was assessed by DAPI (blue). A differential interference contrast (DIC) image is also shown (gray). The tem1Δ::GAL1-UPL-TEM1 strain allows for the rapid, conditional depletion of Tem1. UPL, which stands for ubiquitin-proline-LacI, acts as a destabilizing module that permits rapid degradation of appended proteins. Image is Fig 1B, middle panels, in J Cell Biol. (2011) 192: 599-614. Other images in Fig 1 include CIL #13882, 13883, 13884, 13885, 13886, 13887.

本数据集包含中期细胞（Cell Image Library编号CIL# 13883）与后期细胞（本图像）中Tem1组成型靶向至纺锤体极体（spindle pole body, SPB）的成像数据。实验采用从着丝粒（CEN）质粒中表达增强绿色荧光蛋白（enhanced green fluorescent protein, eGFP）标记的CNM67–TEM1融合蛋白的tem1Δ::GAL-UPL-TEM1菌株，在含2%棉子糖与2%半乳糖的培养基中培养。图像展示了细胞转移至含2%葡萄糖的培养基后，后期细胞中Cnm67-Tem1融合蛋白的定位情况（eGFP荧光信号呈绿色）。Tem1正常情况下优先定位于有丝分裂后期进入子细胞的SPB（对应CIL# 13882、13885）。Cnm67是SPB外斑的整合组分，Cnm67-Tem1融合蛋白的定位在SPB复制完成至有丝分裂结束前均呈对称分布。细胞核形态通过4',6-二脒基-2-苯基吲哚（DAPI，蓝色）染色进行评估。本成像数据同时附带微分干涉差（differential interference contrast, DIC）成像结果（灰度信号）。tem1Δ::GAL1-UPL-TEM1菌株可实现Tem1的快速条件性耗竭。泛素-脯氨酸-LacI（ubiquitin-proline-LacI, UPL）作为去稳定模块，可介导所融合蛋白的快速降解。本图像为《细胞生物学杂志》(2011) 192卷：599-614页中图1B的中间面板。图1其余图像对应的CIL编号为13882、13883、13884、13885、13886、13887。