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NB-LRR immune receptors in plants play dual roles as sentries and as activators of defense. The site in the cell where these activities take place can be different for different NB-LRRs. Furthermore, recognition and defense activation can occur in distinct subcellular compartments. Therefore, determining the subcellular localization of NB-LRRs is a key step toward understanding how they function. Recent advances in confocal microscopy enable high-resolution imaging of proteins in live cells. Agroinfiltration in the
Nicotiana benthamiana
model plant system is a convenient way of expressing proteins for localization studies. This chapter explains how to use
N. benthamiana
to transiently express NB-LRRs for confocal fluorescence microscopy.