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Figure 6 | Plant Methods

Figure 6

From: Screening for in planta protein-protein interactions combining bimolecular fluorescence complementation with flow cytometry

Figure 6

Results of FRET-FLIM measurements of CPK3 with candidate interaction proteins. A. Example expression domains of CPK3-eGFP and mRFP-XP fusion proteins as seen in tobacco epidermal cells. B. Comparison of the localization pattern of APX3 with either an N-terminal mRFP fusion (left) or a C-terminal mCherry fusion (right). The unhampered fusion is the N-terminal mRFP fusion which shows an uneven expression domain unlike the C-terminal mCherry fusion. C. Example of false-colored FRET-FLIM imaging sectors. Stronger FRET-FLIM results in a reduction in the average lifetime. The negative control in CPK3-eGFP alone; the positive control is a CPK3-eGFP-mCherry (called CPK3-FRET) fusion; and real sample of CPK3-eGFP co-expressed with mRFP-ORP2A. D. The averages of the average lifetime (from the measurement sector) are shown as box plots for the CPK3-eGFP in the presence of mRFP-XP fusions. Letters are significance classes based on Students t-test; any sample not connected by a letter is significantly different from the CPK3-eGFP control. Significance difference to the control using Dunnett’s Method is indicated by a *. E. The averages of the average lifetime (from the measurement sector) are shown as box plots for the CPK3-eGFP in the presence of XP-mCherry fusions. Letters are significance classes based on Students t-test; any sample not connected by a letter is significantly different from the CPK3-eGFP control. Significance difference to the control using Dunnett’s Method is indicated by a *. Cytoplasm and Nucleus indicate in which compartment the measurements were made.

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