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Anaphase B data was calculated using GFP-Tubulin images (Ptk2 cells) or GFP-centrin images (HeLa cells) or DIC images (HeLa cells) by measuring the length of the spindle between the two spindle poles.
During their movement towards the ring, a very subtle pattern could be seen: the dots were slightly more prevalent in the region of the central spindle, i.e., in a large diamond-shaped region between the two spindle poles.
We conclude that loss of a single centriole leads to a difference in minus-end microtubule stability between the two spindle poles, that this difference plays a large role in the asymmetric positioning of the metaphase plate, and that cells reduce this difference as they center the metaphase plate before anaphase onset.
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Then, the angle between the vector determined by the two spindle poles and the vector between the centroid of the embryo and the middle of the spindle was calculated using the Numbers software (Apple Computer).
Then, the angle between the vector determined by the two spindle poles (P1P2) and the vector going from the centroid of the embryo to the middle of the spindle (OC) was calculated using the iWorks Numbers software (Apple Computer).
We measured the acute angle between the spindle axis (defined as a line connecting the two spindle poles) and a line extending from the cell body's centroid to the midpoint between the duplicated flagella's basal bodies.
The two spindle poles are connected by a thread of microtubules that form the mitotic spindle.
While the chromosomes attempted to congress within a bipolar spindle, two supernumerary spindle poles appeared under the mCherry-tubulin fluorescence (30 min); thereafter, the four spindle poles further separated and the chromosomes re-organized within a multipolar spindle (50 min).
DOI: http://dx.doi.org/10.7554/eLife.02208.002 The budding yeast spindle consists of ∼40 microtubules (MTs) that extend between the two opposed spindle pole bodies (SPBs).
A direct comparison between Nobs and NMMO for wild-type and cdc25.22 spindles reveals that all but four of the fourteen spindles deviate from the theoretical model by less than a single microtubule, thus indicating precise scaling of microtubule number to changes in spindle length and the availability of polymerised tubulin.
Our measurements revealed that the differences in dissipation of kinetochore-microtubules between the two half-spindles were as high as the differences amongst the k-fibers within a half-spindle, suggesting that any difference between the two half-spindles is likely to be small, and thus outside of the sensitivity range of the photoactivation assay.
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