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At 180 min, DNA integrity was mostly restored in mutation-corrected cells (~90% DNA integrity), whereas the mutant cells still showed significantly reduced (~60%) DNA integrity, confirming that the observed ligation defect and delayed repair are indeed caused by the FUS mutations.
Although reduced, mutant cells still retained staining that localized in a perinuclear manner (Fig. 2B).
Bra mutant cells still undergo an EMT and therefore could be subject to these movements.
Thus, we tested whether the cdc24-4 mutant cells still compartmentalized the ER membrane properly under this regime.
However, mutant cells still expressed other notable hPGC genes, including SOX17, TFAP2C, TEAD4, and OCT4, albeit at lower levels.
Note that shuttle vectors transforming Δ cpaAIR mutant cells still require CpaI methylation, which can be readily performed using Dam+ E. coli strains for plasmid propagation.
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Since the incompletely-segregated dipM mutant cell still produced a certain level of the DipM protein and was still able to divide (doubling time of dipM mutant was similar to that of the wild type), we examined DipM localization in the dipM mutant.
Whereas cilia in CCNO-mutant cells still contain motility-related proteins such as DNAH5 and CCDC39 and can display normal beating patterns, MCIDAS-mutant cells are immotile and lack those axonemal motor proteins [1, 2].
However, the membrane-associated pool of Crb in ept,stat92E/+ mutant cells is still be detected overlapping the Dlg-positive basolateral domain (see arrowhead Fig. 2D-D″).
At this stage, the ph0 mutant cells are still integrated in the follicular epithelium, but they have undergone cell shape changes that indicate segregation from the rest of the ph+ epithelial cells.
The cell size reduction is proportional: PTEN mutant cells are still enlarged with respect to control cells under starvation.
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