Plenti 3 ubc luc2

Lentivirale particle were generated by co-transfection of HEK293T Lenti-X cells (Clontech) with packaging plasmids (pMD2.G, pMDLg/pRRE, pRSV-REV, Addgene) and the lentiviral transfervector (pLenti-III-UbC-Luc2, Applied Biological Materials Inc.) expressing Luciferase 2. After 48 and 72 hours the supernatants were pooled, filtered through a 45 um filter and ultra centrifuged at 32000 rpm 4°C for 1h. The virus titers were determined by HIV p24 antigen test (Elecsys, Roche Diagnostics GmbH). MEB-Med-8A cells were transduced with virus particles (MOI 5) and thereafter selected with puromycin for cell clones that show stable luciferase expression (Invitrogen).

Lentivirale particles were generated by co-transfection of HEK293T Lenti-X cells (Clontech) with packaging plasmids (pMD2.G, pMDLg/pRRE, pRSV-REV, Addgen) and the lentiviral transfervector (pLenti-III-UbC-Luc2, Applied Biological Materials Inc.) expressing Luciferase 2. After 48 and 72 hours the supernatants were pooled, filtered through a 45 um filter and ultracentrifuged at 87076 g 4°C for 2 h. The virus titers were determined by HIV p24 antigen test (Elecsys, Roche Diagnostics). MEB-Med-8A cells were transduced with virus particles (MOI 5) and thereafter selected with puromycin and validated for homogenous luciferase activity (Invitrogen).