Figure 2.
Green fluorescent protein (GFP) expression driven by an nuclear factor of activated T-cells (NFAT) responsive promoter in transduced peripheral blood lymphocytes (PBLs) was triggered by T cell activation. (a) Schematic representation of retroviral vectors: MSGV1.GFP and MSGV1.NFAT.GFP.PA2 GFP, enhanced green fluorescence protein; LTR, long-terminal repeat; NFAT, composite NFAT-responsive promoter element; PA2, polyadenylation signal; SA, splice acceptor; SD, splice donor. (b) Vectors were transfected 293GP cell using lipofectamine 2000 for transient virus production (top histograms). PBLs were transduced with individual vectors as described in the methods. Two days later, the cells were left untreated (middle histograms) or activated with phorbol myristate acetate (PMA)/ionomycin overnight (bottom histograms). Expression of GFP in 293GP cells and PBLs was measured by fluorescence-activated cell sorter (FACS), the percent positive cells was as shown. UT, untransduced.