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We do not believe that Mad3p degradation is critical to turn off the spindle checkpoint, although it is possible that it could play a role in checkpoint adaptation, and this is currently being explored.
An increasing number of kMTs may also switch off the spindle assembly checkpoint in merotelic states (class 4) over time.
The gradual increase of kMTs may help turn off the spindle assembly checkpoint in normal conditions but can promote a faulty conformation (merotelic attachments) to evade the checkpoint.
Although there could be several ways to turn off the spindle checkpoint, we propose that PP1 plays an important role in several of these silencing mechanisms.
Ye et al. propose that, once all chromosomes are lined up on the spindle, TRIP13 turns off the spindle assembly checkpoint by converting closed MAD2 to open MAD2.
Therefore, the predicted gradual increase of kMTs in amphitelic states (class 5) (Fig. 3 a and Additional file 1: Figure S7A) may switch off the spindle assembly checkpoint progressively.
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Thread the nut back on a few turns to keep it from falling completely off of the spindle initially.
Once the cup is removed off of the spindle, turn the frame over and remove the bottom bracket itself, by turning clockwise.
The same effect was also observed at 10 μM nocodazole or 15 μM colchicine, ruling out an off-target effect of the spindle poison on the checkpoint, or an effect of residual microtubules on checkpoint satisfaction (Supplementary Figure S4).
Cytokinesis takes place in the plane through the center of the spindle, so the off-centered spindle apparatus results in an asymmetric cell division.
Once the chromosomes are in place, the checkpoint is turned off and the spindle pulls the chromatids apart so that each daughter cell receives a complete set of chromosomes.
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