In the MFM group, b-tubulin III positive cells concentrated in the border of the neurospheres, while GFAP and Nestin positive cells kept the same localization.. We highlight the fact that Nestin localization in the CTR group was the same as that of BrdU positive cells. Despite Nestin localization was not altered, it is important to notice that the percentage of positive cells significantly decreases, explaining the increased differentiation in the borders of the neurosphere after mitogens removal. b-tubulin III expression increased in hNPC and displayed a clear tendency of increasing in mNPC , after growth factors removal, as showed by real-time PCR.Together, these results reinforce the hypothesis that mitogens removal, even without adhesion and migration, is responsible for an increased cell differentiation. We hypothesize the shift in the distribution of b-tubulin III positive cells was caused by a decreased gradient of growth factors from the outer layer to the center of neurospheres. Given that the concentration of EGF and FGF-2 inside the neurosphere might be lower than in the outside , cells in the neurosphere core are able to stop proliferation and start differentiation even in suspension. DNA microarrays are important experimental tools to gain knowledge about the steady state levels of mRNA species. Affymetrix GeneChips were designed to contain a series of oligonucleotide probes complementary to a specific mRNA of known genes. To quantify a specific mRNA species, the signals from a group of probes representing a specific gene are normalized and averaged. However, often the design of the array and the selection of probe sequences were finalized before the human genome was fully annotated. Therefore some probes lack specificity and the conventional probe sets do not always reflect current knowledge about the multiple individual transcripts encoded by the same gene. Furthermore, mRNA processing mechanisms can lead to different transcripts of the same gene which can have specific biological EX 527 functions. Methods that apply microarray profiling would provide additional information not on the expression levels of a gene but also the respective splice isoforms. This finding is consistent with the observed defect in IkBa phosphorylation and degradation. Therefore, TRAF6 is selectively required for canonical NF-kB activation upon TLR or CD40 stimulation but not for activation of the alternative pathway downstream of CD40 or BAFF-R. Again, these results point towards a role for TRAF6 in B cell subset specification rather than in generating survival signals for homeostasis. The involvement of insulin/IGF1 signalling in lifespan regulation in mammalian species was first suggested in Ames and Snell dwarf mice in which insulin/IGF1 function is reduced due to a deficiency in growth hormone. Recent observations in knockout mice further provided evidence for a direct role of reduced insulin/IGF1 signalling in regulation of mammalian lifespan.