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. 2001 Sep 17;194(6):733–746. doi: 10.1084/jem.194.6.733

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© 2001 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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E47 deficiency promotes the aberrant development of RAG-1 null thymocytes. (A) Dot plots depicting surface CD4 and CD8 expression, as determined by flow cytometry, on thymocytes from 1-mo-old mice with RAG-1^+/− E47^+/−, RAG-1^+/− E47^−/−, RAG-1^−/− E47^−/−, and RAG-1^−/− E4^−/− genotypes. The RAG-1^+/− E47^−/− mouse used for this experiment was from a separate litter and analyzed separately from the other three mice. Numbers indicate the percentage of viable cells found in each quadrant. (B) Flow cytometric analysis of surface CD44 and CD25 expression on CD3⁻CD4⁻CD8⁻ thymocytes from a 3.5-wk-old RAG-1^−/− E47^−/− mouse and three littermates. Numbers indicate the percentage of viable DN cells found in each quadrant. (C) Dot plots depicting surface CD4 and CD8 expression on thymocytes from 1-mo-old RAG-1^−/− littermates that were either wild-type or heterozygous for E47, illustrating the appearance of DP thymocytes in some RAG-1^−/− E47^+/− mice. The percentage of DP thymocytes observed in RAG-1^−/− E47^+/− mice with significant numbers of DP thymocytes has ranged from ∼2–85% (data not shown). (D) Scatter plot indicating the number of thymocytes obtained from six RAG-1^−/− E47^−/− mice as compared with RAG-1^−/− E47 wt or ^+/− littermates. Thymus cellularity in E47^−/− mice typically ranges from 1.5–5 × 10⁷ cells (data not shown). (E) Histograms of surface CD95 (Fas antigen) expression in selected thymocyte populations as measured by flow cytometry. (Thick line) DP thymocytes from a 4-wk-old E47^−/− RAG-1^−/− mouse. (Thin line) DN thymocytes from a RAG-1^−/− E47^+/− littermate. (Dotted line) DP thymocytes from a RAG-1^−/− E47^+/− littermate. (F) Histograms of BCL-2 expression levels in selected thymocyte populations as measured by flow cytometry. (Thick line) DP thymocytes from a 1-mo-old E47^−/− RAG-1^−/− mouse. (Thin line) DN thymocytes from a RAG-1^−/− E47 wild-type littermate. (Dotted line) DP thymocytes from a RAG-1^+/− E47^+/+ littermate. All data presented are representative of analyses of at least three individual RAG-1^−/− E47^−/− mice and littermate controls.

E47 deficiency promotes the aberrant development of RAG-1 null thymocytes. (A) Dot plots depicting surface CD4 and CD8 expression, as determined by flow cytometry, on thymocytes from 1-mo-old mice with RAG-1^+/− E47^+/−, RAG-1^+/− E47^−/−, RAG-1^−/− E47^−/−, and RAG-1^−/− E4^−/− genotypes. The RAG-1^+/− E47^−/− mouse used for this experiment was from a separate litter and analyzed separately from the other three mice. Numbers indicate the percentage of viable cells found in each quadrant. (B) Flow cytometric analysis of surface CD44 and CD25 expression on CD3⁻CD4⁻CD8⁻ thymocytes from a 3.5-wk-old RAG-1^−/− E47^−/− mouse and three littermates. Numbers indicate the percentage of viable DN cells found in each quadrant. (C) Dot plots depicting surface CD4 and CD8 expression on thymocytes from 1-mo-old RAG-1^−/− littermates that were either wild-type or heterozygous for E47, illustrating the appearance of DP thymocytes in some RAG-1^−/− E47^+/− mice. The percentage of DP thymocytes observed in RAG-1^−/− E47^+/− mice with significant numbers of DP thymocytes has ranged from ∼2–85% (data not shown). (D) Scatter plot indicating the number of thymocytes obtained from six RAG-1^−/− E47^−/− mice as compared with RAG-1^−/− E47 wt or ^+/− littermates. Thymus cellularity in E47^−/− mice typically ranges from 1.5–5 × 10⁷ cells (data not shown). (E) Histograms of surface CD95 (Fas antigen) expression in selected thymocyte populations as measured by flow cytometry. (Thick line) DP thymocytes from a 4-wk-old E47^−/− RAG-1^−/− mouse. (Thin line) DN thymocytes from a RAG-1^−/− E47^+/− littermate. (Dotted line) DP thymocytes from a RAG-1^−/− E47^+/− littermate. (F) Histograms of BCL-2 expression levels in selected thymocyte populations as measured by flow cytometry. (Thick line) DP thymocytes from a 1-mo-old E47^−/− RAG-1^−/− mouse. (Thin line) DN thymocytes from a RAG-1^−/− E47 wild-type littermate. (Dotted line) DP thymocytes from a RAG-1^+/− E47^+/+ littermate. All data presented are representative of analyses of at least three individual RAG-1^−/− E47^−/− mice and littermate controls.