Using fluorescence resonance energy transfer (FRET) measured by fluorescence lifetime imaging microscopy (FLIM) we have explored the protein-protein interactions between fluorescent protein tagged fusion proteins of the activation pathways of PKC and NFkB. We observe FRET between CFP-IκB and YFP-p65 in unstimulated cells and when treated with TNFα. We also observed a reduction of the fluorescent lifetime of CFP-IκB in the absence of YFP-p65 when TNFα is present.
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