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Initiation of stress granule assembly by rapid clustering of IGF2BP proteins upon osmotic shock

Biochim Biophys Acta Mol Cell Res. 2020-07-01; 
Wei-Jie Zeng, Chuxin Lu, Yuanyuan Shi, Chunyan Wu, Xinxin Chen, Chunmei Li, Jie Yao
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Custom DNA/RNA Oligos … Cells were hybridized with 70 ng Cy3-labeled 30-mer oligo(dT) probes (synthesized by Genscript, Nanjing, China) overnight at 37 °C. After hybridization, cells were washed once with 4× SSC buffer and then once with 2× SSC buffer … Get A Quote

摘要

Stress granules (SGs) are membraneless organelles formed in the cytoplasm by liquid-liquid phase separation (LLPS) of translationally-stalled mRNA and RNA-binding proteins during stress response. Understanding the mechanisms governing SG assembly requires imaging SG formation in real time. Although numerous SG proteins have been identified, the kinetics of their recruitment during SG assembly has not been well established. Here we used live cell imaging and super-resolution imaging to visualize SG assembly in human cells. We found that IGF2BP proteins formed microscopically visible clusters in living cells almost instantaneously after osmotic stress, followed by fusion of clusters and the recruitment of G3BP1 a... More

关键词

Cell imaging, Osmotic stress, Phase separation, Stress granule, mRNA