Intracellular Staining by Flow Cytometry
|Detection of Autoimmune Regulator/AIRE in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes were stained with Rat Anti-Mouse Autoimmune Regulator/AIRE APC‑conjugated Monoclonal Antibody (Catalog # IC6184A) and Rat Anti-CD11c PE‑conjugated Monoclonal Antibody (Catalog # FAB69501P). Quadrant markers were set based on control antibody staining (Catalog # IC006A). View our protocol for Staining Intracellular Molecules.|
Autoimmune Regulator (AIRE) is an approximately 60 kDa nuclear and cytosolic protein that is required for the development of T cell tolerance. It regulates the expression of self-antigens by both thymic epithelial cells and extrathymic cell types. Genes known to be regulated by AIRE include FABP2, Proinsulin, Oxytocin, MCP-2, S100A8 and Cytochrome p450. The genes, referred to as TRAs, represent non-thymic, tissue-specific molecules that could serve as autoantigens at a later date. Their expression in the thymus (and select extrthymic sites) allows for the indentification and elimination of potentially autoreactive T cell clones. While some mouse and human molecules are identical in their regulation by AIRE, possibly not all are, and this leads to subtle differences in the autoimmune syndromes seen in human and mouse that are attributable to AIRE absence or mutation. AIRE regulates gene transcription through interactions with DNA, histone H3, and the nuclear matrix. It contains one HSR domain that is involved in dimerization (aa 1-106), a nuclear localization sequence (aa 114-134), one SAND domain involved in DNA binding (aa 182-282), and two PHD zinc finger domains that mediate protein-protein interactions (aa 298-345 and aa 433-475). Alternate splicing of mouse AIRE generates at least eleven isoforms. Within aa 476-552, mouse AIRE shares 91% and 64% aa sequence identity with rat and human AIRE, respectively.
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