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36% �� 4.07% [http://en.wikipedia.org/wiki/Bumetanide Bumetanide] of GFP+ EV cells and 43% �� 4.8% of GFP+ WT cells; mean �� SD, n?= three independent transplant experiments; p?[http://www.selleckchem.com/products/cobimetinib-gdc-0973-rg7420.html MEK inhibitor] from the first round of transplantation because this can have deleterious effects on transplant potential. In the first round of transplants, as previously, no differences in the number of EV, S155R, or WT outgrowths were observed, suggesting that there was no effect on initial engraftment. Again, however, the S155R outgrowths were less extensive. In the second round of transplants, whereas both the EV and WT-expressing cells engrafted successfully and with equal potency, only a single rudimentary S155R outgrowth formed (Figure?3K), suggesting that the self-renewal of mammary stem cells expressing S155R in the primary graft had been inhibited. Overall, therefore, these data supported a model in which WT AURKA activity enriched for cells with a luminal progenitor phenotype, whereas expression of mutant S155R AURKA promoted the basal myoepithelial cell fate while suppressing stem cell self-renewal. Next, we addressed potential mechanisms by which AURKA may regulate mammary epithelial cell fate and differentiation. [http://www.selleckchem.com/products/MLN8237.html Alisertib research buy] In Drosophila neuroblasts, AURKA regulates asymmetrical divisions and cell fate by promoting alignment of the mitotic spindle with the cortical polarity axis ( Lee et?al., 2006). We therefore investigated a role for AURKA in regulating the orientation of the mitotic spindle in the cap cells of TEBs. We examined mitotic spindle orientations in cap cells of the unmanipulated developing pubertal mammary gland (Figure?4A) and in the invading ductal ends of outgrowths developing from transplanted mammary epithelial cells transduced with EV, WT (Figures 4B and 4C), or S155R lentiviruses.