Histological Analysis of SLC38A6 (SNAT6) Expression in Mouse Brain Shows Selective Expression in Excitatory Neurons with High Expression in the Synapses
Figure 6
Proximity ligation assay (PLA) on N25/2 hypothalamic cell line to demonstrate protein-protein interaction pattern between SLC38A6 and neuronal markers.
The nuclei of the cells are stained in blue with DAPI. The PLA signals are detected with red filter. A) SLC38A6 interacts with Snap25 exhibiting proximity to the membrane. B) SLC38A6 interacts with Synaptotagmin, illustrating substantial proximity to the vesicular proteins. C) PLA control with no primary antibody incubation and hence no signal of interaction between proteins demonstrating successful PLA in the treated cells. D) Signals per cell were counted and represented here as histograms. Columns 1, 2, 3, 4 and 5 reveal interactions between Synaptotagmin-Synaptotagmin, Snap25-Snap25, Synaptotagmin-Snap25, SLC38A6-Synaptotagmin and SLC38A6-Snap25, respectively. Columns 1 and 2 exhibit quantification of PLA signals obtained by using different antibodies on the same proteins, Synaptotagmin and Snap25 respectively. Column 3 depicts interaction between synaptotagmin and Snap25 to illustrate their proximity in the cells. Columns 4 and 5 show interactions between SLC38A6 and synaptotagmin and Snap25, respectively. E) Normalized signals per cell are shown here to compare interactions of SLC38A6 with Synaptotagmin and Snap25, respectively. Column 1 and column 2 demonstrate normalized interactions between SLC38A6 and synaptotagmin and SLC38A6 and Snap25, respectively.