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Protocol for screening anti-amyloidogenic compounds in cultured mammalian cells
Emma Lacroix1, Sandra Keerthisinghe2, Evgenia A Momchilova1
1Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, British Columbia V5A 1S6, Canada; Centre for Cell Biology, Development, and Disease, Simon Fraser University, Burnaby, British Columbia V5A 1S6, Canada.
None:
A functional form of amyloid aggregate can be rapidly induced by various environmental stressors, yielding subnuclear structures called amyloid bodies (A-bodies). Here, we describe a cell-based high-throughput assay to identify anti-amyloidogenic compounds capable of impairing heat-induced A-body formation in adherent cell lines. We describe steps for generating reporter cell lines, optimizing scoring algorithms, screening compound libraries, and analyzing results. This assay can be expanded to various systems to identify compounds with both anti- and pro-amyloidogenic effects.
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