Figure 6: The effect of three different point mutations within the SARAH domain of RASSF5C on the dimerisation with the isolated SARAHMST1.

(A) Plate map showing the average EGFP lifetimes calculated for 10 fields of view per well fitted to a monoexponential decay model. (B) Box plots showing median EGFP lifetimes, interquartile (box range), standard deviation (whisker), 1% and 99% percentile (×) and minimum/maximum values (−) for the segmented cells in different conditions within the plate: green: EGFP-RASSF5C (wild type and mutants) only; red: EGFP-RASSF5C (wild type and mutants) + mCherry-SARAHMST1; blue: EGFP-RASSF5C (wild type and mutants) + mCherry (see Supplementary material for a table of differences in mean fluorescence lifetime). (C) Average intensity ratios acceptor/donor (ImCherry/IEGFP) for the segmented cells in different conditions within the plate (same colour code as in B). (D) 2D plots of intensity ratios acceptor/donor 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.