Additional file 1.

Figure S1. Isolation and characterization of CD45-UEA1+ mTECs and CD45-UEA1- thymic stromal cells. Isolation and characterization of CD45-UEA1+ mTECs and CD45-UEA1- thymic stromal cells. A. Isolation of CD45-UEA1+ mTECs (red) and CD45-UEA- thymic stromal cells (yellow) by flow cytometry. Values are representatives of the percentage of each population in repeated isolations. B. Gene expression in mTECs, thymic stromal cells, thymus, liver and ES cells by RT-PCR. Compared with the stromal cells, higher expressions of Aire, Cd80, Krt5, Krt14, Krt8, Foxn1 in both the Aire+/+ and Aire−/− mTECs were detected. The Aire−/− mice strain used in this research lacks the exon2 of Aire, but the transcription of Aire could be detected [1]. E2, exon2; E3, exon3; E5, exon5. C. Real-time RT-PCR quantification of transcript levels of Aire in mTECs isolated from Aire+/+ mice, as well as mTEC, thymus from C57BL/6 strain, and ES cells. The Exon2 to Exon3 of Aire mRNA was amplified. Standard curve method calculation was used to compute transcript levels relative to Actb mRNA. The y axis indicates arbitrary unit normalized to Actb, with the expression level in thymus as 1. The expression level of Aire in mTECs was over 50 times higher than in the thymus, which showed a similar pattern as human AIRE in mTECs and the thymus [2]. The expression level of Aire in ES was about half of the thymus. Expression of Aire in liver was not detected.

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Wu et al. BMC Immunology 2012 13:58   doi:10.1186/1471-2172-13-58