Figure 10

GluN2 and GFP transgene expression in vitro mediated by lentivirus. In this experiment, 293FT cells were transduced with lentiviruses designed to express GFP, Flag-GluN2A, or Flag-GluN2B from a CMV or a TRE3G promoter as indicated. TRE3G promoter containing viruses were transduced into cells that were also transfected with the pTet-Off plasmid. Forty eight hours after viral transduction, native GFP expression was observed via fluorescence microscopy and Flag-GluN2 expression was observed via ICC and fluorescence microscopy. Images depict DAPI stained nuclei with the same fields viewed for GFP or Flag-GluN2 (Texas Red) transgene expression. Non-transduced cells and cells transfected with pRK5-Flag-GluN2 plasmids were processed as negative and positive ICC controls respectively. These viruses were capable of conferring GluN2 or GFP transgene expression as intended.