Fig. 1. Crp and Ins2 promoters drive model antigen expression in distinct patterns among mTEC subsets.

a Tg mouse constructs. b Relative USA expression in sorted mTEC^lo and mTEC^hi from Aire^+/+ or Aire^−/− CRP and INS2 Tg mice. c USA mRNA expression in mTEC^lo and mTEC^hi from CRP, CRP^lo, INS2 and INS2^lo Tg mice. d Calculation of USA mRNA expression in total mTEC. The thymus from three mice were pooled together prior to sorting, and data are representative of at least two independent experiments. e–i Single-cell RNA sequencing of total thymic epithelial cells (TEC) from CRP, CRP^lo, and INS2 Tg mice. e Uniform Manifold Approximation and Projection (UMAP) of TEC from CRP Tg mice. f UMAP highlighting Crp, Ins2, and USA transgene expression in TEC from CRP Tg mice. g Distribution of the Crp- or USA-expressing cells among the different TEC clusters isolated from the three Tg mice (Crp) or from CRP Tg mice (USA). h Proportion of mTEC^lo and mTEC^hi cells that express Crp or Ins2 from the three Tg mice. i Average log-normalized per cell expression of Crp and Ins2 among mTEC^lo and mTEC^hi isolated from the three Tg mice. Data from g–i are from one experiment with one mouse per Tg line and are represented as mean ± SEM for analysis of Crp and Ins2. j Representative histograms of mature OT-I T cell proliferation and activation 72 h after overlay on the indicated thymic slices as indicated by CFSE dilution as well as CD44 and PD-1 expression. k Compilation of the proportion of CFSE^lo OT-I T cells 72 h after overlay on the indicated thymic slices (mean ± SEM, n = 12 LMC, 23 CRP, 15 CRP^lo, 24 INS2, 15 INS2^lo, from 3 independent experiments). l Compilation of the mean fluorescence intensity (MFI) of CD44 and PD-1 and m the proliferation index of the proliferating mature OT-I T cells. Littermate control (LMC) (mean ± SEM, n = 3 LMC, 17 CRP, 6 CRP^lo, 18 INS2, 6 INS2^lo, from 3 independent experiments). Source data are provided as a Source Data file.