Direct Assessment of the Effect of the Gly380Arg Achondroplasia Mutation on FGFR3 Dimerization Using Quantitative Imaging FRET
Figure 1
Donor, FRET and acceptor scans of a single vesicle.
One vesicle co-expressing the two wild-type constructs tagged with eYFP (donor) and mCherry (acceptor) is imaged in three different scans: the donor (ex: 488 nm, em: 500–530 nm), FRET (ex:488 nm, em: 565–615 nm) and acceptor (ex: 543 nm, em: 650 nm long-pass) scans. The intensities across the membranes are obtained by summing along the vesicle circumference. They are shown as a function of the distance from the vesicle center (blue). After background correction, they are fitted with Gaussians (green) and the Gaussians are integrated to yield total channel intensities. The red curve is the difference between the data and the fit.