Figure 4: Comparison of the RASSF family members in terms of dimerisation with the SARAHMST1 domain using FRET. | Scientific Reports

Figure 4: Comparison of the RASSF family members in terms of dimerisation with the SARAHMST1 domain using FRET.

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

Figure 4

(A) Plate map showing average EGFP donor lifetimes (ps) calculated for 10 fields of view (FOV) per well using a monoexponential fit. (B) False-colour FLIM images of cells from a typical FOV in each well showing the EGFP lifetime (ps) per pixel. (C) Box plots showing median EGFP lifetimes, interquartile (box range), standard deviation (whisker), 1% and 99% percentile (×) and minimum/maximum values (−) calculated for individual cells averaged over 10 FOV per well using monoexponential analysis: green: EGFP-RASSF(1–10) only; red: EGFP-RASSF(1–10) + mCherry-SARAHMST1; blue: EGFP-RASSF(1–10) + mCherry-MST1ΔSARAH (see Supplementary material for a table of differences in mean fluorescence lifetime). (D) Acceptor/donor intensity ratios (ImCherry/IEGFP) averaged over each cell for all the conditions in the plate. The colour code is the same as in C). (E) Scattered plots of EGFP lifetimes versus acceptor/donor intensity ratios (ImCherry/IEGFP) calculated for individual cells (with same colour code as for C). FLIM data were acquired with wide-field imaging.

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