Sequence diversity among proliferating and nonproliferating Vβ2-Jβ1.1 and Vβ4-Jβ1.1 CD44⁺, Foxp3⁻ cells CD4 T cells (1).

<p>Experimental approach to analyze the CDR3 sequences of Vβ2-Jβ1.1 and Vβ4-Jβ1.1 CD4⁺ CD44⁺ Foxp3⁻, KI-67^bright and Ki-67^negative cells from individual mice. Cells were FACS sorted into lysate buffer containing proteinase K. One-third of the lysate was subjected to PCR with specific Vβ and Jβ1.1 primers (see Methods). PCR products were cloned into the Topo vector followed by bacterial transformation and single colony isolation. PCR with universal M13 primers were used on each colony to amplify the Vβ2-Jβ1.1 and Vβ4-Jβ1.1 gene segments followed by sequencing with the universal T3 primer.