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Herein, we observed that the tubes obtained with Penetratin and CF-Penetratin were thinner in PC GUV compared to those observed in PC/PG GUV (Fig 2A D).
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As expected:[ 8, 25] 1) the shift in wavelengths and GP values between ordered and disordered phase is much less pronounced than in GUVs (ordered: GP≈0.2 versus ≈0.4 in GUVs, and disordered: GP≈−0.15 versus ≈−0.5 in GUVs, compare Figures 2 and 3),[ 17] and 2) the lipid packing in non-phase-separated GPMVs (GP≈−0.05) is in-between that of the Lo and Ld environments of the phase-separated GPMVs.
Equilibrium dialysis showed (a) different permeation of CPFX through the membranes of GUVs and multilamellar vesicles (MLVs), with characteristic half-lives of 54.4 and 18.1 h, respectively; and (b) increased retention of CPFX in case of GUVs with released amounts of 70% compared with about 97% in case of MLVs.
The π π interactions between aromatic rings are corroborated by our observation of a redshifted fluorescence of B12 in the star-shaped domains of DPPC GUVs, when visualized with a spectral detector, compared with B12 dissolved in organic solvent.
Importantly, full-length Lsp1 displays similar dynamics in vivo compared to its isolated BAR domain on GUVs, suggesting that this protein efficiently also diminishes the lateral diffusion of lipids at the plasma membrane of living cells.
However, time-lapse imaging of giant unilamellar vesicles (GUVs) in the presence of the protein domains suggested that the heterodimeric Rvs161/167 BAR domain is slightly more efficient in tubulating membranes compared to the two endocytic F-BAR domains.
Our approach is therefore more general compared to a recently published approach, which is based on phasor analysis of spectral images recorded for Laurdan on a two-photon microscope.[ 18] We first exemplified GP spectral imaging on model membranes such as giant unilamellar vesicles (GUVs) composed of different mixtures of lipids.
This becomes even more obvious when comparing the distribution of GP values over images of GUVs from the direct sampling (non-fitting) with that of the curve-fitting analysis.
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