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Depletion of Wdr8 results in shorter, tilted spindle microtubules.
This tilted spindle phenotype is circumstantially linked to the depletion of Pax6-positive aNPs (Fig. 4; see Discussion).
Consistently, upon conducting the microtubule regrowth assay in mitotic cells, we did not see any defects in hMsd1-depleted cells during the initial stage of microtubule nucleation around the centrosome; however after 30 min regrowth, tilted spindle microtubules with compromised astral microtubules were formed (supplementary Fig S4G and H).
Furthermore, aNPs in E15 neocortices (mostly RGCs) depleted of p600 or expressing the Ndel1-binding p600 fragment exhibit tilted spindle (Fig. 2B, Fig. 3D) and premature neuronal differentiation, as evidenced by the decreased number of Pax6-positive aNPs and increased number of Tbr2-positive basal progenitors destined to become neurons (Fig. 4).
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Cytokinesis in plants often begins with tilted spindles, indicating the absence of a spindle alignment checkpoint in plant cells.
It is worth noting that the defective phenotypes of shorter and tilted spindles were very similar, if not identical, to those induced by the deletion of hMsd1/SSX2IP [6].
We previously showed that hMsd1/SSX2IP is also required for spindle anchoring and its depletion led to the emergence of shorter, tilted spindles accompanied with dissociation of astral microtubules from the mitotic centrosome.
In sum, NESCs in p600SC−/− brains have tilted mitotic spindle.
These Pax6-positive aNPs display a tilted mitotic spindle.
To assess the physiological consequence of MGRN1-mediated polyubiquitination of α-tubulin, tilt in spindle poles was used as a read-out.
Nevertheless, these newborn Tis21 GFP-positivee BPs, generaTis21 GFP-positive the spindle, may still temporarily exhiBPs certain AP features, such as an AP-like cell cycle lengeneratedh is shorter than that of mature bys (Arai et al., 2011).
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