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For calculations of the number of Aire + and ΔNp63 + TECs, sections were scanned as described above for Aire, ΔNp63 and UEA1.
For Aire and FoxP3 intracellular stain, surface staining was performed first, followed by fixation and permeabilization utilizing the buffers in the Foxp3 Staining Set (eBioscience) according to the manufacturer's instructions.
HSR is central for AIRE di- or oligomerization [4,5].
Immature mTECs are negative for AIRE and CD80 expression, which are activated when the cells differentiate into mature mTECs.
So far, the only known protein partner for AIRE is a general transcriptional co-regulator and histone acetyltransferase, CREBP binding protein (CBP).
Previous work showed that lymphotoxin signaling is required for Aire and Aire-dependent as well as Aire-independent TRA expression [ 52].
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We and others have recently shown that the PHD finger of AIRE interacts with the unmodified N-terminal ends of histone H3, and this interaction is required for AIRE-dependent transcriptional activation (11– 11).
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Thus, these cells have all the necessary factors for AIRE-mediated gene activation and provide a feasible model for studying the molecular mechanisms of ectopic expression.
In contrast, analysis of 154 genes indicated that their downregulation was not dependent on intact PHD1 structure (Fig. 1C and D; Supplementary Material, Fig. S1B), suggesting that histone binding is not essential for AIRE-mediated gene repression.
Dry as for Airing above.
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