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Percentage of BrdU positive cells in the APS treatment group was lower than that in the control group (P<0.01, Figure 1B).
More recently, we developed a BrdU label-retaining assay that permits direct evaluation of stem cell number within each PS and showed that this assay can be used to assess stem cell self-renewal activity (Hu et al.
In the last 6 hours of culture, stimulated cells were exposed to bromodeoxyuridine (BrdU) (BD Pharmingen(tm) - FITC BrdU Flow Kit).
Cell cultures were incubated with 0.24 [micro]M 5'-bromo-2'-deoxyuridine (BrdU) for approximately two cell cycles.
PCNA was only found at the outside of the microencapsulated cell spheres, but not in the center (Figure 6(d)), which was a similar trend to BrdU shown above.
In addition, BrdU was used to detect cell proliferation, and the percent of BrdU-positive cells in the control group (30.24% [+ or -] 0.89%) was significantly higher than that in the CSE group (19.49% [+ or -] 0.62%) (p < 0.05; Supplementary Figure 3C).
Data were represented as a number of positive cells/[mm.sup.2] of BrdU or double-positive cells per section.
E2 and ER agonists significantly suppressed cell proliferation, indicated by lowered BrdU incorporation, compared with the vehicle-treated cells (Figure 1(b)).
For histologic examination, the frozen sections were subjected to hematoxylin-eosin staining, immunostaining for BrdU, p63, and CK (cytokeratin) 3/12.
To determine whether homeostatic proliferation occurred in vivo, we treated mice with BrdU and collected spleens to detect for BrdU incorporation.
Immunohistochemistry reactions were performed using antibodies against Ki67 (Dako, Glostrup, Denmark; clone MIB-1, DAKO 7240, dilution 1:200); BrdU (Neomarkers, Fremont, CA, USA; clone Bu20a,1848-P, dilution 1:1000); BCL2 (Dako M0887, clone 124, dilution 1:500); [p27.sup.Kip1] (Dako M7203, clone Sx53G8, dilution 1:1000); Vitamin D receptor (Millipore, Temecula, CA, USA; clone 9A7, Millipore 04-1526, dilution 1:30), followed by detection through Lab Vision[TM] UltraVision[TM] LP Detection System: HRP Polymer/DAB Plus Chromogen (visualized using 3.3'diaminobenzedine tetrahydrochloride (DAB, D-5637, Sigma, St.