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. 2023 May 24;15:1156777. doi: 10.3389/fnsyn.2023.1156777

FIGURE 3.

FIGURE 3

GluN1, GluN2A, and GluN3A subunit proteins colocalize for making t-NMDARs in the MEA. (A) Quadruple immunolabeling of GluN1 (images in R1-4C1), GluN2A (images in R1-4C2), and GluN3A (images in R1-4C3) subunit protein puncta relative to dendrites immunolabeled with MAP2 in the MEA (red box in the bottom left inset of image in R1C1) and their pairwise and triplicate colocalization (merged images in R2-3C4, R5C1-4) for making t-NMDARs (top left inset in R1C1) at the indicated enlargements (lettered boxes in yellow). Arrowheads point to representative examples of individual subunit puncta or their colocalization. Changes in the pseudocolor antigen key (marked by || between images) are to aid in gauging colocalization of the proteins imaged. (B) The merge matrix for pairwise assessment of GluN1, GluN2A, and GluN3A subunit protein colocalization at a single synapse on a dendrite at the highest level of magnification. The merged images in the rightmost column (R1-2C4) showcase t-NMDAR subunit composition, comprising glycine binding subunits (GluN1 and GluN3A) coalescing with glutamate binding subunits (GluN2A).