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Proliferating cells are hardly detectable in the adult mammalian brain by microscopy of stained sections, but after pre-labeling with radioactive thymidine or 5′-bromo-2-deoxyuridine (BrdU), either marks the nucleus, as do mitosis-related proteins such as Ki67 and PCNA.
DAPI (blue) marks the nucleus.
Indeed, Importin-β bound the NTE under cytoplasmic conditions and became disengaged by RanGTP, which marks the nucleus as the destination compartment of import (Görlich et al, 1996).
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In some cases 1 μg/ml DAPI was added to live cells immediately prior to imaging to mark the nucleus.
Tissue sections (4 µm) were prepared and counterstained with neutral red marking the nucleus and sometimes the cytoplasm at a lower level.
The RFP was attached to a chromatin protein to mark the nucleus and the RFP value was used to normalise the YFP and CFP values on a per-cell basis to control against imaging variations.
The number of nuclei per dorsal tracheal branch was counted using live embryos expressing btl-GAL4/UAS-α-Cat to mark the junctions and UAS-NLS-red-stinger to mark the nucleus, excluding the branches at the very anterior and posterior of each embryo.
To mark the nuclei of transfected cells after extraction, cells were also co-transfected with a plasmid encoding a known chromatin-bound protein GFP-MCM7.
Hoechst stain was used to mark the nuclei.
Detection was performed with 3′3′diaminobenzidine (Dako, Denmark) and counterstained with hematoxylin for marking the nuclei.
The lymphatic vessels were immunostained in cryosections with anti-Prox1 antibodies, which mark the nuclei of LECs.
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