Tracking protein aggregation and mislocalization in cells with flow cytometry
Yasmin M Ramdzan, Saskia Polling, Cheryl PZ Chia, Ivan HW Ng, Angelique R Ormsby, Nathan P Croft, Anthony W Purcell, Marie A Bogoyevitch, Dominic CH Ng, Paul A Gleeson, Danny M Hatters
NATURE METHODS | NATURE PUBLISHING GROUP | Published : 2012
We applied pulse-shape analysis (PulSA) to monitor protein localization changes in mammalian cells by flow cytometry. PulSA enabled high-throughput tracking of protein aggregation, translocation from the cytoplasm to the nucleus and trafficking from the plasma membrane to the Golgi as well as stress-granule formation. Combining PulSA with tetracysteine-based oligomer sensors in a cell model of Huntington's disease enabled further separation of cells enriched with monomers, oligomers and inclusion bodies.
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Awarded by Australian National Health and Medical Research Council
This work was funded by grants to D. M. H. (Australian National Health and Medical Research Council project grant 566640). D. M. H. is funded as a Grimwade fellow, supported by the Miegunyah Trust. D.C.H.N. is funded as a CR Roper fellow. The SOD1-GFP fusion constructs were provided by B.J. Turner (Florey Neurosciences Institute, Australia). HSPA1A (Hsp70) and DNAJB1 (Hsp40) in the pCMV vector was provided by P. Muchowski (Gladstone Institute of Neurological Disease and University of California, San Francisco).