American Society of Hematology

Fig. 4.

$Fig. 4. B220lo. / c-kit+CD19− cells retain their TCRβ locus in a germline configuration but express T-lineage–specific genes. (A) DNA was prepared from 5 × 103 cells from fraction e before and after FTOC. Rearrangement of the TCRβ genes was examined by a sensitive 2-step PCR using pairs of primers (1 and 2) followed by a runoff using the fluorescent primer 3, as shown in the higher panel. TCRβ gene rearrangement was not observed in cells from fraction e. In contrast, diverse Dβ-Jβ rearrangements (indicated by arrows) were observed after FTOC. (B) RT-PCR analysis of cells from population e (line 2) for the indicated genes. Controls (line 3) included 15-dpc fetal thymocytes, CD19+B220+ cells, and NK1.1+CD3− mature NK cells. S17 cells were used as a negative control (line 1). The amount of cDNA was carefully standardized according to HPRT transcripts.$

B220^lo

c-kit⁺CD19⁻ cells retain their TCRβ locus in a germline configuration but express T-lineage–specific genes. (A) DNA was prepared from 5 × 10³ cells from fraction e before and after FTOC. Rearrangement of the TCRβ genes was examined by a sensitive 2-step PCR using pairs of primers (1 and 2) followed by a runoff using the fluorescent primer 3, as shown in the higher panel. TCRβ gene rearrangement was not observed in cells from fraction e. In contrast, diverse Dβ-Jβ rearrangements (indicated by arrows) were observed after FTOC. (B) RT-PCR analysis of cells from population e (line 2) for the indicated genes. Controls (line 3) included 15-dpc fetal thymocytes, CD19⁺B220⁺ cells, and NK1.1⁺CD3⁻ mature NK cells. S17 cells were used as a negative control (line 1). The amount of cDNA was carefully standardized according to HPRT transcripts.

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