FIGURE 6.

Surface expressed GluN2C co-localizes with synaptophysin, a presynaptic marker. A, hippocampal neurons were transfected with GFP-GluN2C or GFP-GluN2A. To visualize the surface receptors, cells were incubated with anti-GFP antibody, fixed and incubated with Alexa 647-conjugated anti-rabbit secondary antibody (depicted in red for optimal visual contrast effect). Cells were then permeabilized and labeled with anti-synaptophysin antibody and Alexa 568-conjugated anti-mouse secondary antibody (depicted in green), a pre-synaptic vesicle protein marker. The right panels display representative zoomed in higher magnification images of 10 μm dendritic segments. Scale bar = 10 μm. B, co-localization of GFP puncta to synaptophysin puncta was measured. Value is given in percent co-localization (defined as the number of GFP puncta overlapping or adjacent to synaptophysin puncta, divided by the total number of pre-synaptic puncta). Data represent means ± S.E. (n = 12 cells per group; n = 36 dendritic segments per group; Student's t test, *, p < 0.05).