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A Protocol for Using Förster Resonance Energy Transfer (FRET)-force Biosensors to Measure Mechanical Forces across the Nuclear LINC Complex
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Chapters
- 00:05Title
- 00:53Transfect Cells with Nesprin-2G and Other Plasmid DNA
- 04:09Verify Transfection Efficiency
- 05:12Capture Spectral Fingerprints of mTFP1 and Venus Fluorophores for Spectral Unmixing
- 06:53Capture Unmixed Images
- 07:52Results: Setting Imaging Parameters and Capturing Spectrally Resolved FRET Images
- 08:51Conclusion
A number of FRET-based force biosensors have recently been developed, enabling the protein-specific resolution of intracellular force. In this protocol, we demonstrate how one of these sensors, designed for the linker of the nucleoskeleton-cytoskeleton (LINC) complex protein Nesprin-2G can be used to measure actomyosin forces on the nuclear LINC complex.
