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. 2013 Feb 11;210(2):287–300. doi: 10.1084/jem.20122149

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© 2013 Aichinger et al.

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Figure 9. — Autophagy-dependent direct presentation by mTECs is essential for negative selection at limiting antigen expression. (a) Expression of GCL mRNA in thymic stromal cells from Aire-GCL or Aire-GCL^lo mice as determined by qPCR. Values (mean ± SD from three independent biological replicates) are normalized to expression in Aire-GCL mTEC^hi. (b) GFP fluorescence in mTECs from WT (gray) and Aire-GCL^lo (black line) mice. Data are representative of three independent experiments with pooled cells from four or more animals per genotype. (c) Thymi of DEP single-tg, Aire-GCL × DEP, and Aire-GCL^lo × DEP mice were analyzed by flow cytometry. The mean frequency ± SD of CD4SP cells (top row) and of DEP-TCR⁺ cells (Vα11⁺Vβ5.1⁺) among gated CD4SP thymocytes (bottom row) is indicated. (d) Frequencies and absolute numbers of CD4SPVα11⁺Vβ5.1⁺ cells in mice of the indicated genotypes. Data in c and d are from ≥8 mice per genotype. (e) Thymocyte subsets in Aire-GCL^lo × Atg5^+/− or Aire-GCL^lo × Atg5^−/− thymic lobes 4–6 wk after transplantation into DEP × MHC Class II^+/+ recipients. The mean frequency ± SD of CD4SP cells (top row) and of DEP-TCR⁺ cells (Vα11⁺Vβ5.1⁺) among gated CD4SP thymocytes (bottom row) is indicated. Data are representative of at least 17 chimeras per group. (f) Frequencies and absolute numbers of CD4SP Vα11⁺Vβ5.1⁺ cells among total thymocytes in (e) as compared with antigen nonexpressing (WT) Atg5^+/− (n = 19) or Atg5^−/− (n = 20) thymic lobes. Data from antigen nonexpressing thymi are also shown in Fig. 8 a.