Skip to main content
. 2016 Jan 5;110(1):176–187. doi: 10.1016/j.bpj.2015.11.032

Figure 5.

Figure 5

Characterization of membrane fluidity in competition assays. Measurements of the lateral diffusion constants of fluorescently labeled probe lipids preinserted into GUVs or delivered by interacting LUVs, using time-lapse images from FRAP experiments. (a) Selected fluorescence image sequences of NBD-labeled POPC GUVs (green) and material decorating the GUV surface delivered by Rho-B-labeled POPC LUVs (red) from the surrounding bulk. Fluorescence was monitored before and directly after bleaching, until complete recovery was achieved. (i–iii) GUV fluorescent probe (green) recovery in (i) the absence of LUVs or AH peptide (prebleach, 0 s, 1 s, 2 s, 100 s after bleaching; Movie S4), (ii) the presence of LUVs without incubation with AH peptide (prebleach, 0 s, 1 s, 2 s, 60 s; Movie S15), and (iii) the presence of LUVs and after incubation with 2 μM AH peptide (prebleach, 0 s, 2 s, 5 s, 50 s; Movie S16). (iv) LUV-derived material with Rho-B (red) decorating the GUV, showing recovery after addition of and incubation with 2 μM AH peptide (prebleach, 0 s, 2 s, 5 s, 60 s; Movie S17). Scale bars, 10 μm. (b–e) Normalized fluorescence recovery curves for FRAP experiments: (b) (e-i), D = 9.56 μm2/s; (c) (e-ii), D = 9.38 μm2/s; (d) (e-iii), D = 2.15 μm2/s; and (e) (e-iv), D = 1.83 μm2/s. To see this figure in color, go online.