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Figure 5 | Plant Methods

Figure 5

From: The FAST technique: a simplified Agrobacterium-based transformation method for transient gene expression analysis in seedlings of Arabidopsis and other plant species

Figure 5

FRET and BiFC evidence for HY5 homodimerization using the FAST assays. Arabidopsis seedlings were cocultivated with agrobacteria cells simultaneously carrying two FRET or BiFC constructs except for (B). A, The negative FRET control, soluble YFP and Cerulean alone, showed a low Nfret (normalized FRET) value in a representative region of the cotyledon. Note blue color. Scale bar = 60 μm. B, The positive FRET control, a YFP-Cerulean fusion, showed a high Nfret value in a representative region of the cotyledon. Note green to yellow color. The remaining Cerulean signal was barely visible due to the intensive energy transfer from Cerulean to YFP. Scale bar = 60 μm. C, YFP-HY5 and Cerulean-HY5 showed a medium Nfret value in a representative region of the cotyledon. Note light blue to green color. Scale bar = 60 μm. D, Quantification of the average Nfret value in a large number of cells for each FRET combination indicated. E, Reconstituted YFP fluorescence in a BiFC combination of YN-HY5 and YC-HY5 in a representative region of the cotyledon. Scale bar = 60 μm.

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