Generation of NKT-ES and NKT-ES-V cells by direct nuclear transfer of mature NKT cells. (A) Schematic representation of the TCRα locus in natural killer T embryonic stem (NKT-ES) and NKT-ES-V cells. The NKT-ES cells generated by nuclear transfer of B6 liver NKT cells were prepared as described elsewhere,^27,28  with a slight modification. NKT-ES-V cells were generated by transfection of the Cα-IRES-Venus targeting vector, which placed the Venus cDNA, along with its upstream IRES, downstream of TCRα locus adjacent to the Cα stop codon. (B) Genomic polymerase chain reaction (PCR) of NKT-ES and NKT-ES-V cells. Primer pairs shown in panel A were used for genomic PCR. Primer sequences are listed in supplemental Table 2. Lane 4 is clone 3, which was used in the present study.