Figure 6.

HA-MC4R localizes together with and in the proximity of GluN2A. (A) Confocal images (20 µm scale bar) and SR images (5 µm scale bar) of an HA-MC4R-expressing neuron, immunostained with mouse anti-HA Tag (F-7) and anti-NR2A (N327/95) to visualize endogenous GluN2A. The white rectangle indicates the relative region of the enlarged images in the panel below. The white arrow indicates a point of colocalization, and the cyan/magenta arrows indicate a site where GluN2A and HA-MC4R are adjacent. (B) Enlarged images of the neurite above where HA-MC4R and GluN2A colocalize (white arrow) and where they are adjacent (cyan/magenta arrow). The yellow dashed lines indicate those drawn for the segment analyses, with the corresponding graphs displayed as means +/− SEM on the right. (C) The distance (nm) between HA-MC4R and GluN2A fluorescence peaks was determined by line segment analyses from 5 neurons from 2 independent experiments (n = 39 colocalizing points, n = 19 adjacent points). The green dotted line in the graph is as in Figure 1. Models indicate the range of distances between colocalizing and adjacent peaks of HA-MC4R and GluN2A fluorescence, respectively.