Figure 2.

Total lipid mixing assay of inner membrane vesicles (IMVs). (A) Fluorescence emission spectra of IMVs labelled with NBD-PC (donor, D) and RhoPE (acceptor, A) at a different molar ratios. The excitation wavelength was 460 nm. The peaks at 523 and 593 nm correspond to the maximum fluorescence emission of the donor and acceptor respectively. An optimal Förster resonance energy transfer (FRET) from NBD to Rhodamine was observed at molar ratios of 1:5 between the donor and the acceptor. (B) FRET efficiency ratio of IMVs (left), Mfn1-IMVs (middle) and E. coli total lipid extract (TLE) large unilamellar vesicles (LUVs) (right), before and after 60 min incubation with 10 mM GTP. Student’s t-test was performed to measure the significance of statistical difference between the different groups. $p\le 0.05,(\ast )$ and $p\le 0.001,(\ast \ast \ast )$ were considered statistically significant.