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Markers for mitotic dysregulation, such as supernumerary centrosomes and altered centrosome-related mRNA and protein expression, were observed in gliomas as compared to normal tissue.
We have now used anti-tubulin immunostaining of mitotic microtubule arrays and DAPI staining of nuclear chromatin as reliable markers for mitotic staging of dividing cells.
The experiment has been repeated, using anti-tubulin immunostaining of mitotic microtubule arrays and DAPI staining of nuclear chromatin as reliable markers for mitotic staging of dividing cells.
Markers for mitotic dysregulation, such as supernumerary centrosomes and altered expression of centrosome-related mRNA and proteins were more frequently detected in higher grade gliomas.
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Double staining for DE-cad and phospho-histone H3 (a marker for mitotic chromosomes) showed that these cells were not mitotic (data not shown) suggesting the AJ defects may be associated with neuroblast delamination.
Phosphorylated Histone 3 used as a marker for mitotic cells.
Acetylated tubulin was used as a marker for mitotic spindle poles.
Next, cells were fluorescently labeled for DNA (DAPI staining) and phosphorylated histone H3 (pH3), a marker for mitotic cells.
Thus, the phosphorylation of histone H3 at serine 10 (pSer10) is used as a sensitive marker for mitotic cells (18).
Proliferation was also assessed in the OCRL1 mutant embryos using phospho-histone H3 (PH3) as a marker for mitotic cells.
Mitotic arrest was confirmed by the presence of Histone 3 phosphorylation (P-H3), a marker for mitotic chromosome condensation (Hendzel et al., 1997).
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