The Journal of Neuroscience, December 8, 2004, ():
Interactions of Postsynaptic Density-95 and the NMDA Receptor 2 Subunit Control Calpain-Mediated Cleavage of the NMDA Receptor
J. Neurosci. Dong et al.
24: 11035
Supplemental data
Files in this Data Supplement:
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Supplemental Figure 1. Clustering of NR2A and PSD-95 in transfected cells. HEK293 cells were transfected with the NR1/NR2A combination in addition to wildtype (WT) PSD-95 or PSD-95 (C3,5S) mutant. Using specific antibodies that recognize PSD-95 (green) and NR2A (red), we visualized the cellular localization of PSD-95 comparing WT and mutant forms by confocal microscopy. WT PSD-95 fluorescence was consistently visualized in clusters (first row with PSD-95 shown in first column). In addition, PSD-95 clusters localized with NR2A clusters at the cell surface (NR2A shown in second column, and merged image shown in last column). PSD-95 (C3,5S) mutant (second row) was visualized diffusely throughout the cytoplasm with no note of clusters. Colocalization of PSD-95 (C3,5S) with NR2A fluorescence appeared coincidental since there was no predominant fluorescence of PSD-95 (C3,5S) in any portion of the cytoplasm. Therefore, the PSD-95 (C3,5S) mutant, which abolishes palmitoylation, also prevents clustering of NMDA receptors as demonstrated previously (El-Husseini et al., 2000).
