Figure 5: Effect of three different point mutations within the SARAH domain of RASSF1 on the dimerisation with the isolated SARAHMST1. | Scientific Reports

Figure 5: Effect of three different point mutations within the SARAH domain of RASSF1 on the dimerisation with the isolated SARAHMST1.

From: Screening for protein-protein interactions using Förster resonance energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM)

Figure 5

(A) Plate map showing the average EGFP lifetimes calculated for 10 fields of view per well when fitting to a monoexponential decay profile. The wild-type EGFP-RASSF1 assay shows that mCherry alone can serve as a negative control as well as the mCherry-MST1ΔSARAH. (B) Box plots showing median EGFP lifetimes, interquartile (box range), standard deviation (whisker), 1% and 99% percentile (×) and minimum/maximum values (−) for segmented cells in different conditions within the plate: green: EGFP-RASSF1 (wild type and mutants) only; red: EGFP-RASSF1 (wild type and mutants) + mCherry-SARAHMST1; blue: EGFP-RASSF1 (wild type and mutants) + mCherry (see Supplementary material for a table of differences in mean fluorescence lifetime). (C) Average acceptor/donor intensity ratios (ImCherry/IEGFP) for the segmented cells in different conditions within the plate (same colour code as in B). (D) 2D plots of acceptor/donor intensity ratios versus EGFP lifetimes for the segmented cells in different conditions within the plate (same colour code as in B). FLIM data were acquired with wide-field imaging.

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