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Surprisingly, we found that Kog1 and Tco89 move from their known location on the vacuolar membrane (marked with Vph1-mCherry), to a single body near the edge of the vacuole, within 60 min in most cells.
Regions in the bottom part of the plasma membrane (marked with a green square) showed a high mobile fraction similar to untreated cells, indicating that the peptide was not able to diffuse into the gap between the cell and the glass slide.
When comparing T24 cells incubated with 10 μM KLK to untreated cells, two effects became apparent: Firstly, the plasma membrane showed dramatic morphological changes including numerous membrane invaginations (marked with white arrows), and additional circular structures near the plasma membrane (marked with gray arrows) (Fig. 8A C).
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The band corresponding to cleaved PINK1 on the Coomassie-stained membrane was marked with an asterisk before the membrane was desiccated using silica gel overnight.
LC3-II accumulated primarily in the crude lysosomal/heavy membrane fraction (marked with LAMP1, Supplementary Figure S1a and c), confirming translocation to the membrane.
The CP110 binding partner Cep97 also remained associated with mother centrioles docked at the plasma membrane, marked by CD44.
CP110 labeling appeared closer than centrin-3 to the plasma membrane (marked by CD8 or PKC-θ) in 95% CTLs with docked centrosomes (n = 55), indicating that CP110 was not lost from centrioles on association with the plasma membrane.
Leading edge membrane marked by dashed box is enlarged below.
Numerous evidently maturing AGJ/autophagosome-intermediates, with double-membrane cisternae (marked with arrows) progressively encircling AGJs, were clearly identifiable (Fig. 3B, panels a c).
Over the next three minutes, the membrane of the phagosome marked with a circle loses VatM-GFP, apparently through vesiculation.
The orange colour dots that appeared within the cells are possibly the outcome of a combination between the red dyed membrane and the phage marked with green colour generated by the secondary FITC conjugated antibody, which may suggest a lysosomal incorporation of the internalized phages.
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CEO of Professional Science Editing for Scientists @ prosciediting.com