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Fig. 13 | Plant Methods

Fig. 13

From: One pattern analysis (OPA) for the quantitative determination of protein interactions in plant cells

Fig. 13

No detectable interaction between MADS-domain proteins with FRET-FLIM in Arabidopsis GFP/mCh reporter. FLIM experiments were performed in stable A. thaliana lines. MADS-box proteins fused with the indicated FP were expressed by their endogenous promoter. Donor only images were acquired in homozygous GFP reporter lines. For the FRET samples, GFP lines were crossed with the respective mCherry reporter and FLIM images were acquired in resulting F1 plants. Each data point was calculated from a respective FLIM image containing several nuclei. Only nuclei containing both donor and acceptor were considered in FLIM image analysis of FRET samples. BINDING [%] (grey) and FRET efficiencies [%] (white) for SEP3-GFP, SEP3-GFP AP1-mCh, AP3-GFP, AP3-GFP PI-mCh, AP3-GFP AP1-mCh, PI-GFP and PI-GFP AP1-mCh. No increased BINDING above the 10% cut-off in FRET samples was detectable for any of the tested combinations. Statistical groups were assigned after multiple comparison with Kruskal–Wallis and a Post hoc test using the criterium Fisher’s least significant difference (alpha parameter is 0.05) (Dashed blue line marks the BINDING Cut-Off of 10%; Number of repetitions are indicated below BINDING values and number of images with BINDING above 10% are indicated below the FRET efficiency values in the bottom of the plot)

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