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It remains controversial whether traditional endothelial-derived capillaries also service these tumor cells, with opinions based on the stain used, the observer, and the intratumoral location of the observation.
The PicoGreen stain exclusion assay differentiates between membrane-enclosed and non-enclosed DNA based on the stain's inability to cross a lipid bilayer.
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'Fluorescence minus one (FMO)' controls based on the staining combination used for the experimental sample but in which one antibody was left out and replaced with its isotype control were also used to set sort gates correctly.
Quadrants were generated based on the staining of isotype antibodies so that isotype control- stained cells were in the bottom part of the quadrant.
The immunostaining score (IS) was based on the staining intensity among these categories and on the percentage of stained cells.
Similar to the in vitro study, the cytoplasmic presence of most of NF-κB was observed in the untreated control (Fig. 4Bi), and nuclear translocation was induced by IL-1β based on the stain-filled nuclei (Fig. 4Bii).
The optimum ratio was experimentally determined based on the staining signals (8 µM peptide in the Figure 6).
The "percent reconstitution" shown in figures was based on the staining results of anti-CD45.2 and anti-CD45.1.
Based on the staining intensity, percentages of differentiated, half-differentiated and undifferentiated cells were determined by counting.
The analysis was based on the staining pattern of the Cot-1 RNA fraction (hsRNA), which decorates transcriptionally active areas of DNA, using Cot-1 probe in non-denaturing conditions [33].
The evaluation was based on the staining intensity and extent of staining.
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