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. 2015 Feb 24;8:12. doi: 10.1186/s13041-015-0100-7

Figure 9.

Figure 9

Lentiviral plasmids confer spurious transgene expression in vitro , due to the presence of the RSV promoter at the 5′ end of the lentiviral genome. In this experiment, AAV and lentiviral plasmids designed to express Flag-GluN2A/B from a 0.4αCaMKII promoter, and a lentiviral plasmid designed to express Flag-GluN2A from a 0.4αCaMKII promoter which had the RSV promoter deleted (−RSV), were transfected into 293FT cells. Twenty four hours after transfection Flag-GluN2 expression was observed via ICC and fluorescence microscopy. Images depict DAPI stained nuclei with the same fields viewed Flag-GluN2 (Texas Red) transgene expression. Similar to previous results, the AAV plasmids did not confer GluN2 expression and the lentiviral plasmids did confer GluN2 transgene expression. However the lentiviral plasmid without the RSV promoter did not confer 0.4αCaMKII-GluN2A transgene expression. Untransfected cells and cells transfected with pRK5-Flag-GluN2 plasmids were processed as negative and positive ICC controls respectively, (scale bar = 20 μm).