T Cell Receptor δ Gene Rearrangement and T Early α (TEA) Expression in Immature αβ LineageThymocytes: Implications for αβ/γδ Lineage Commitment  Anne Wilson, Jean-Pierre de Villartay, H.Robson MacDonald  Immunity  Volume 4, Issue 1, Pages 37-45 (January 1996) DOI: 10.1016/S1074-7613(00)80296-4

Figure 1 TCR Gene Expression in Subsets of Immature and Mature T Cells Thymocyte subsets and lymph node T cells were prepared as described in Experimental Procedures. Both CD25+ and CD25− DN subsets were sorted to be CD3loHSA+CD44lo. Total RNA (4 μg) was loaded into each lane. The same blot was sequentially hybridized with the indicated probes. Exposure times were as follows: 48 hr for Cα and Cδ; 18 hr for Cβ; 24 hr for Cγ; and 6 hr for β-actin. TCR β transcripts in SP cells, which are expressed at much lower levels than in DP thymocytes (Maguire et al. 1990), could readily be detected in longer exposures (data not shown). Immunity 1996 4, 37-45DOI: (10.1016/S1074-7613(00)80296-4)

Figure 2 Expression of TEA in Thymus Subsets Subsets and controls are as in Figure 1. Exposure times were 2 weeks for TEA and overnight for β-actin. Immunity 1996 4, 37-45DOI: (10.1016/S1074-7613(00)80296-4)

Figure 3 FACS Sorting of ISP Thymocytes CD4−CD25− thymocytes were stained with anti-CD8–Red613, anti-HSA–PE, and anti-CD3–fluorescein isothiocyanate and the CD8+CD3loHSAhi (ISP) cells sorted. The sorting gate is indicated and on reanalysis after sorting the cells were >98% pure. Immunity 1996 4, 37-45DOI: (10.1016/S1074-7613(00)80296-4)

Figure 4 TCR δ and TEA Expression by ISP Thymocyte Hybridomas Total cell RNA was prepared and 5 μg loaded into each lane. Hybridization was performed sequentially with the TEA probe, the Cδ probe, and then the β-actin probe. Exposure times were as follows: 5 days for TEA; overnight for Cδ; 6 hr for β-actin. Immunity 1996 4, 37-45DOI: (10.1016/S1074-7613(00)80296-4)

Figure 5 Expression of TCR δ Transcripts in Mature αβ T Cells from TCR Transgenic Mice Total RNA from lymph node cells of αβ transgenic or littermate control mice was blotted and sequentially probed with Cδ and Cα. The longer TCR α transcript originates from the transgenic cDNA construct (Pircher et al. 1989) Ethidium bromide staining is indicated as a measure of RNA loading. Immunity 1996 4, 37-45DOI: (10.1016/S1074-7613(00)80296-4)