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. Author manuscript; available in PMC: 2009 May 26.
Published in final edited form as: J Neurosci. 2008 Nov 26;28(48):12604–12613. doi: 10.1523/JNEUROSCI.2958-08.2008

Figure 5. PSDs recovered after RAP-induced reversal of Tat-induced synapse loss contain NMDA receptor immunoreactivity.

Figure 5

A, Representative images display labeled PSDs on a neuron expressing PSD95-GFP and DsRed2 before (0 h) and during (16 & 24 h) treatment with 50 ng/ml Tat. The LRP inhibitor RAP (50 nM) was added after 16 h. 8 hr after administering RAP the number of synapses recovered (24 hr frame). B, After collecting the live cell image at 24 hr (as shown in A 24 h frame) the cells were fixed and labeled with antibodies to DsRed and the NR2A and 2B subunits of the NMDA receptor as described in Methods. Confocal micrographs display PSD95-GFP fluorescence (green) and NR2A and NR2B (red) immunoreactivity. Merged images display overlapping puncta (yellow). Processed images display puncta within the DsRed2 mask (blue). The same image-processing algorithm described in Methods was used to identify both NR2 immunoreactive puncta and PSD95-GFP puncta. The insets are enlarged images of the boxed region. Note that NR2A and NR2B immunoreactivity includes non-transfected cells in the field. Scale bar represents 10 µm.