Extended Data Fig. 5: Exclusive expression of I-A^g7-WT in bone marrow derived cells allows the development of tetramer^bright IA^g7-ChgA_HIP reactive T cells in I-A^g7-WT/WT and I-A^g7-PD/WT hosts.

(a) Representative staining from concatenated data (n = 2 mice) of CD4SP thymocytes from radiation chimeras of I-A^g7-WT/WT bone marrow (BM) into I-A^g7-WT/WT, I-A^g7-PD/WT, I-A^g7-PD/PD or H2^b expressing mice. (b) Overlay of I-A^g7-ChgA_HIP tetramer⁺ CD4SP thymoctes from I-A^g7-WT/WT and IA^g7-PD/WT chimeras with I-A^g7-WT/WT BM. (c,d) Quantification of the (c) number and (d) percentage of I-A^g7-ChgA_HIP tetramer^bright and tetramer^dim cells among individual I-A^g7-WT/WT BM chimeric mice. (e) Overlay and (f,g) MFI of I-A^g7 staining on cortical and medullarly thymic epithelial cells of 6 weeks old I-A^g7-WT/WT (n = 5), I-A^g7-PD/WT (n = 4), I-A^g7-PD/PD (n = 5) mice. (h–j) Example and quantification of IL-2 release from BDC2.5 T cells hybridomas with co-cultured with ChgA_HIP peptide and sorted (i) cTECs and (j) mTECs populations from I-A^g7-WT/WT, I-A^g7-PD/WT, I-A^g7-PD/PD mice. Example (k) contour plot and (l) plotted average % CTV^dim BDC2.5 CD4 T cells proliferating in response to BM-DCs derived from I-A^g7-WT/WT and I-A^g7-PD/PD presenting β-islet cell preparations based on from 3 replicate wells per experiment using separate islet preparation per experiment. (m,n) Quantification of IL-2 release by BDC2.5 and BDC6.9 T cell hybridomas in response to BM-DCs derived from I-A^g7-WT/WT and I-A^g7-PD/PD presenting β-islet cell preparations or soluble peptide. Bars are mean values from the 6 wells. P values are from a one-way ANOVA with Dunnett’s multiple comparisons test.