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Maximum nuclear staining > 10% (any intensity) was considered positive, as employed in most studies using this antigen.
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The expression of the AR was scored as the percentage of nuclear staining in a maximum of 1000 cells per sample.
Note that pHH3 staining pattern depends on the phosphorylation state of the histone H3 protein, which changes during the cell cycle; only cells with uniformly intense nuclear staining (that reaches a maximum at metaphase) were included in the analyses.
To detect individual cells in tissue sections, a nuclear cell quantification image analysis algorithm (IHC Nuclear Quantification, Aperio) was trained on control slides by defining the color vectors for the hematoxylin nuclear counterstain and primary positive chromagen DAB, minimum and maximum size for nuclei, and threshold ranges for intensity of nuclear staining.
Lastly, DAPI was used for nuclear staining.
Bar, 30 μm. (D) DAPI nuclear staining of morula embryos.
(D) Nuclear staining with DAPI of embryos at E7.5.
Hollow arrows show DAPI-positive nuclear staining (blue).
Apoptosis was measured by nuclear staining and Annexin V-PI staining.
Furthermore, all cases, including those harboring dysplasia, showed negative nuclear staining for p53 and positive nuclear staining for retinoblastoma (RB).
In sporadic adenomas, the glandular architecture is regular, p53 nuclear staining is negative, there is usually strong nuclear staining for beta catenin and positive Bcl2 staining.
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