Transfect Cells with Nesprin-2G and Other Plasmid DNA
4:09
Verify Transfection Efficiency
5:12
Capture Spectral Fingerprints of mTFP1 and Venus Fluorophores for Spectral Unmixing
6:53
Capture Unmixed Images
7:52
Results: Setting Imaging Parameters and Capturing Spectrally Resolved FRET Images
8:51
Conclusion
副本
The overall goal of this procedure is to determine forces on the linker of the nucleoskeleton cytoskeleton, or LINC complex, through FRET microscopy in living cells via a biosensor designed for the LINC complex protein, nesprin-2G. This method can
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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.