Figure 5.
CPK3 Co-Localization and Interaction with TPK1 In Vivo.
(A) Subcellular localization studies of TPK1::GFP (green) in the vacuolar membrane and CPK3 as RFP-fusion protein (red) in the cytoplasm of onion epidermal cells. Co-localization of both proteins is indicated by the yellow sphere close to the vacuolar membrane in the overlays. The lower left images in both panels represent bright field images while lower right images represent a merge of TPK1–GFP and CKP–RFP confocal images. Bimolecular fluorescence complementation of N-terminal fusions of CPK3 and TPK1 (upper panel) or KAB1 (lower panel) with N- and C-terminal parts of CFP, respectively, in tobacco epidermal leaf cells 2 d after infiltration.
(C) Western blot analysis of expression levels shows comparable signal intensities of TPK1 and KAB1 under the different experimental conditions employed to quantify BiFC efficiency.