pMSCV has been described previously (Gilmore et al., 2003 (link)). pMSCV-RELΔTAD1 was created by subcloning a BglII to XhoI fragment containing the RELΔTAD1 cDNA into pMSCV.
Human A293T cells and BJAB or Daudi lymphoma cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10 or 20% fetal bovine serum (Biologos, Montgomery, IL), respectively, as described (Starczynowski et al., 2005 (link)). Virus stocks were generated by transfecting A293T cells with pMSCV or pMSCV-RELΔTAD1 plus helper plasmid pcL10a1, essentially as described previously (Gilmore et al., 2003 (link)). Approximately two days later, virus was harvested. One ml of virus (in the presence of 4 μg/ml polybrene) was used to infect 106 BJAB or Daudi cells using the spin infection method (Gilmore et al., 2003 (link)). Two days later, cells were selected with 2.5 μg/ml puromycin (Sigma) for 2-4 weeks.
Human A293T cells and BJAB or Daudi lymphoma cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10 or 20% fetal bovine serum (Biologos, Montgomery, IL), respectively, as described (Starczynowski et al., 2005 (link)). Virus stocks were generated by transfecting A293T cells with pMSCV or pMSCV-RELΔTAD1 plus helper plasmid pcL10a1, essentially as described previously (Gilmore et al., 2003 (link)). Approximately two days later, virus was harvested. One ml of virus (in the presence of 4 μg/ml polybrene) was used to infect 106 BJAB or Daudi cells using the spin infection method (Gilmore et al., 2003 (link)). Two days later, cells were selected with 2.5 μg/ml puromycin (Sigma) for 2-4 weeks.
