Observing the Assembly of Protein Complexes in Living Eukaryotic Cells in Super-Resolution Using refSOFI
Few approaches are currently available that allow the detection of protein-protein interactions (PPIs) in super-resolution, and the observation of the assembly of protein complexes in living cells has been particularly challenging. We developed reconstituted fluorescence-based stochastic optical fluctuation imaging (refSOFI), which is based on bimolecular fluorescence complementation (BiFC) and SOFI, allowing us to detect protein complex assembly 30 min after the induction of complex formation. Here we describe how to use refSOFI to map the assembly of two proteins of interest into a complex within living cells at super-resolution.
Key wordsSuper-resolution imaging Stochastic optical fluctuation imaging (SOFI) Protein complexes Protein-protein interactions Bimolecular fluorescence complementation (BiFC)
This work was supported by NIH (R35 CA197622, R01 DK073368, R01 GM111665, and R01 MH111516 to J.Z.).
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