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A scratch in the cell monolayer induced directed migration and polarized microtubule networks in cells adjacent to the open scratch zone (the region devoid of cells).
As previously reported, glial cell cultures transduced with Lv-shVIM alone had an intermediate phenotype, one and two weeks after scratching, characterized by lower levels of cellular invasion in the scratch zone and strong residual GFAP immunoreactivity.
By contrast decreasing TREM2 expression correlated with increased numbers of BV-2 cells within in the scratch zone.
Quantification of the number of cells entering the scratch zone was determined based on blinded analysis of digital photographs.
Using cloned BV-2 microglial cell lines, we found that increasing TREM2 levels was associated with a decrease in the number of cells found within the scratch zone 6 h post-scratch.
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Three separate regions along the wound were randomly chosen as scratch zones.
Where control cell (absence of GNP-NKCT1) showed growth in wound zone within 24 h, the treated cell showed very little growth in scratching zone after 48 h.
Phase contrast microscopic images of each scratched zone were taken at a designated time during the 30 h culture period.
Cells treated with AICAR showed enhanced cell migration into the scratched zone, which was attenuated if AMPK, AKT2, or ATF2 were knocked down.
The typical images of cell migration across the scratched zone are shown in Figure 4A, and the number of cells that migrated during the 30 h culture period has been compared in Figure 4B.
Cell migration index (CMI) was defined as [ Nt2 - N0) - (Nt1 - N0)]/ t2 - t1), where Nt2 and Nt1 are the number of cells detected in the scratched zone at 2 different culture times, and N0 is the number of cells at 0 h.
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