FIGURE 7.

Immunofluorescence staining of overexpressed tagged GluN2 subunits. (A–D) Co-transfection of mCherry (mCherry fluorescence) (A–D) and EYFP-tagged GluN2B-encoding plasmids stained with mouse anti-GFP/Alexa-488 (A’–D’). (E–H): Co-transfection of mCherry (mCherry fluorescence) (E–H) and EGFP-tagged GluN2A-encoding plasmids stained with mouse anti-GFP/Alexa-488 (E’–H’). Note intense mCherry staining of immature as well as more mature neurons and the barely visible GluN2A and GluN2B signal. Intensity of the GluN2A and GluN2B immunofluorescence staining increases in the pictures arranged from left to right, and the right-most cells [the pyramidal neuron in (D,D’), and the astrocyte in (H,H’)] had in fact been the two cells we found most intensely labeled for the subunits. Note that the tag fluorescence appeared rather cytosolic, also occluding the nucleus. Double-labeled cells marked by arrows, mCherry-positive cells without GluN2 fluorescence marked by white circles. About 10 OTC transfected from two preparations for each GluN2 subunit; sequential confocal imaging. Pial surface is to the top for all pictures. Scale: 10 μm.