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. Author manuscript; available in PMC: 2011 Jul 15.
Published in final edited form as: J Immunol. 2010 Jun 18;185(2):1055–1062. doi: 10.4049/jimmunol.0903099

Figure 4. Endogenous Vβ Segments Rearrange in Vβ14 NT/+ and Vβ8Tg Thymocytes.

Figure 4

(a) PCR strategy for amplification of Vβ rearrangements to DJβ1.1/DJβ1.2 and DJβ2.1/DJβ2.2 complexes. Shown is a schematic representation of the wild-type TCRβ locus depicting the relative locations of representative Vβ, Dβ, Jβ, and Cβ gene segments, as well as the primers located just downstream of Jβ1.2 or Jβ2.2 and the Cβ2 primers. (b) PCR analysis of potential Vβ-to-DJβ1 and Vβ-to-DJβ2 rearrangements. Shown are representative PCR amplifications of Vβ rearrangements to (top panel) DJβ1.1/DJβ1.2 and (bottom panel) DJβ2.1/DJβ2.2 complexes for the indicated Vβ segments performed on DNA isolated from Vβ8Tg, Vβ14NT/+, or WT thymocytes. The amounts of DNA and numbers of PCR cycles used were previously demonstrated to amplify rearrangements within the linear range for the wild-type sample. (c) Quantitative PCR analysis of Vβ-to-DJβ1 and Vβ-to-DJβ2 rearrangements. Shown are representative PCR amplifications of Vβ rearrangements to DJβ1.1/DJβ1.2 and DJβ2.1/DJβ2.2 complexes for the indicated Vβ segments using serial 1:5 dilutions of DNA isolated from WT, Vβ8Tg, Vβ14NT/+, or Vβ14NT/NT thymocytes. Also shown is a representative PCR amplification of Cβ2 as a control for DNA content.