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. 1998 Oct 19;188(8):1401–1412. doi: 10.1084/jem.188.8.1401

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Intracytoplasmic TCRβ chain expression and DNA content analysis of TCRα/β⁻ thymocyte subsets. (A) Flow cytometry analysis of CD3 surface expression (top panels) or intracytoplasmic (ic) TCRβ (βF1) expression (middle panels) of CD4⁺CD8⁻CD3⁻, large DP CD3⁻, large DP CD3^lowpTα⁺, and small DP CD3⁻ thymocytes isolated as described in Materials and Methods (shaded histograms). Unshaded histograms, Background fluorescence. Cells from each subset were analyzed for DNA content by flow cytometry after staining with PI. Doublet discrimination ensured that only single cells were counted. Percentages of cells in G₀/G₁, S, and G₂/M are indicated (bottom panels). (B) Two-color flow cytometry analysis of TCRβ chain expression versus DNA content of large DP CD3⁻ thymocytes. Cells were electronically gated based on intracytoplasmic (ic) TCRβ chain expression (I, TCRβ⁻; II, TCRβ⁺) and reanalyzed for DNA content.

Intracytoplasmic TCRβ chain expression and DNA content analysis of TCRα/β⁻ thymocyte subsets. (A) Flow cytometry analysis of CD3 surface expression (top panels) or intracytoplasmic (ic) TCRβ (βF1) expression (middle panels) of CD4⁺CD8⁻CD3⁻, large DP CD3⁻, large DP CD3^lowpTα⁺, and small DP CD3⁻ thymocytes isolated as described in Materials and Methods (shaded histograms). Unshaded histograms, Background fluorescence. Cells from each subset were analyzed for DNA content by flow cytometry after staining with PI. Doublet discrimination ensured that only single cells were counted. Percentages of cells in G₀/G₁, S, and G₂/M are indicated (bottom panels). (B) Two-color flow cytometry analysis of TCRβ chain expression versus DNA content of large DP CD3⁻ thymocytes. Cells were electronically gated based on intracytoplasmic (ic) TCRβ chain expression (I, TCRβ⁻; II, TCRβ⁺) and reanalyzed for DNA content.