Figure 1

Adeno-associated viral vector maps and GluN2 and GFP transgene expression from these AAV viral vector plasmids in vitro . (A) Viral genome maps for pAAV-Basic vector backbones containing the following transgenes: Flag-GluN2A/B, Flag-GluN2A/B with C-terminal deletions (GluN2A/BΔC) and GFP. These transgenes are controlled by the 0.4αCaMKII promoter or CMV promoter as indicated. Each of these vectors contain a ~200 bps 3′UTR that contains an SV40 based poly-adenylation signal. GFP = green fluorescence protein, ITR = inverted terminal repeat, MCS = multiple cloning site (B) AAV plasmids designed to express, Flag-GluN2A/B, GFP, and Flag-GluN2A/BΔC from a 0.4αCaMKII promoter and Flag-GluN2A/BΔC from a CMV promoter were transfected into N2A cells. Twenty four hours after transfection, 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. Flag-GluN2A/B did not exhibit detectable expression. However GFP and Flag-GluN2A/BΔC exhibited convincing expression. Untransfected cells and cells transfected with pRK5-Flag-GluN2 plasmids were processed as negative and positive ICC controls respectively. (Scale bar = 20 μm).