Figure 6. Jβ1.1 RS sequences influence transcription and accessibility.

A. Northern blot analysis of germline probe D (see Fig. 3C) hybridizing germline transcripts from Jβ1^σ/σ:Rag-2^-/-, Jβ1^σ.1/σ.1:Rag-2^-/- and Jβ1^σ.2/σ.2:Rag-2^-/- thymocytes. Expression is normalized to GAPDH. B. PCR analysis of Vβ-Dβ rearrangement on the Jβ1^σ.1 and Jβ1^σ.2 alleles in Jβ1^ω/σ.1 and Jβ1^ω/σ.2 thymocytes. The schematic illustrates the PCR strategy using a Vβ-specific primer (PV) and a primer between Dβ1 and Jβ1.1 (P1). PCR analyses were carried out on serial 3-fold dilutions of thymocyte DNA. The PA200 gene is amplified for a DNA loading control. Shown is a representative experiment of three Jβ1^ω/σ.1 and three Jβ1^ω/σ.2 mice analyzed. That the VDβ PCR products were from the Jβ1^σ.1 and Jβ1^σ.2 alleles was confirmed by restriction digestion using unique restriction sites introduced during the targeting.