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This caused a 30% reduction in fork number, but no significant effect on cell cycle distribution or viability.
Furthermore, a short 1 h exposure of NALM-6 cells to 1 μℳ JQ1, insufficient to cause c-Myc downregulation, also led to a marked reduction in fork progression rates.
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We propose that sharpening of the replication profiles upon mrc1 deletion is explained by the associated reduction in fork-velocity.
A reduction in the FORK species compared to the wild-type case suggests a slow-down in S-phase because fewer origins are firing within the prescribed time.
In addition, we observed a severe reduction in the fork velocity upon treatment of NIH3T3 cells with SAHA that inhibits Hdacs1, 2 and 3 (Additional file 11: Figure S11).
We found a consistent reduction in the replication fork velocity in the absence of SMARCA5, which was further exaggerated in the presence of hydroxyurea.
Using genetic knockdown systems and novel Hdacs1,2-selective inhibitors, we found that loss of Hdacs1,2 leads to a reduction in the replication fork velocity, and an increase in replication stress response culminating in DNA damage.
This suggests that a >95% loss of MCM2-7 does not produce a comparable reduction in active replication forks.
It further suggests that S2 cells can tolerate a reduction in the number of active replication forks without a significant effect on the cell cycle distribution or cell viability.
Failure to stabilize and restart stalled forks or prolonged arrest of replication forks (replication stress) may result in fork collapse, leading to chromosomal breakage and rearrangement.
CPT treatment causes replication fork breakage, leading to the reduction in the amount of intact chromosomes that migrated into the gel in wild-type and all swi3 mutant cells.
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