Plasma membrane localization of CAN1 and CAN2 nucleases in protoplasts. ECFP alone and ECFP-fusion proteins were transiently expressed in Arabidopsis root cell protoplasts. Localization was analyzed by confocal laser scanning microscopy (right half of each panel). The corresponding bright-field images are shown on the left halves of panels. (A) Protoplast transformed with empty vector (pSAT6A-ECFP) as a control shows characteristic cytoplasmatic and nuclear localization (B) Expression of the CAN1-ECFP fusion construct. (C) Expression of the CAN2-ECFP fusion construct. Clear plasma membrane localization of both nucleases is seen. The length of the white scale bar at the right bottom of each panel corresponds to 10 μm.