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. Author manuscript; available in PMC: 2018 Dec 1.
Published in final edited form as: Mol Cell Neurosci. 2017 Oct 21;85:190–201. doi: 10.1016/j.mcn.2017.10.004

Figure 3. Overexpression of Gpc5 or Lrrtm4 restores the Rem2 RNAi-dependent decrease in synapse density.

Figure 3

A) Immunostaining for PSD-95 (red) and Synapsin I (blue) in the dendrites of transfected neurons. A synapse is defined as the overlap of red and blue puncta (shown in white) on a GFP positive dendrite. Scale bar = 5µm. B) Quantification of PSD-95/Synapsin puncta for neurons transfected with GFP and either: (Top) a pSuper empty vector (Control), shRNA targeting Rem2 (Rem2 RNAi), Gpc5 cDNA (Gpc5 OE), or shRNA targeting Rem2 and Gpc5 cDNA (Rem2 RNAi + Gpc5 OE) and (Bottom) a pSuper empty vector (Control), shRNA targeting Rem2 (Rem2 RNAi), Lrrtm4 cDNA (Lrrtm4 OE), or shRNA targeting Rem2 and Lrrtm4 cDNA (Rem2 RNAi + Lrrtm4 OE). Data is presented as mean of normalized PSD-95/Synapsin puncta density ± SEM. * indicates p < 0.005 and # indicates P < 0.085 between conditions indicated by bars using two-way ANOVA with Tukey post-hoc test. In both top and bottom graphs, control versus Rem2 RNAi was different at P < 0.005 but this was not indicated on the graph for clarity. Normalized values are the average of ≥ 2 biological replicates and n ≥ 32 neurons for each condition.