Figure S4.
Specific ECM and excitatory synapses at the AIS of AcD and nonAcD neurons. (A and B) Representative images of AIS-specific ECMs labeled by brevican staining at the AIS of DIV21 nonAcD and AcD neurons. Black arrowheads indicate brevican at the AIS. Scale bar is 10 µm. (C and D) Representative images of nonAcD and AcD neurons stained with markers for pre- and postsynaptic compartments of an excitatory synapse (VGLUT and homer-1, respectively), for AIS (TRIM46 or AnkG), and for somato-dendritic compartment (MAP2). (C) VGLUT (D) homer-1. Scale bar is 10 µm. Inset: corresponding zoom-ins of white dashed rectangular. Scale bar is 5 µm. Black arrowheads indicate corresponding markers for pre- and postexcitatory synapse. (E) Distance between co-localized gephyrin and VGAT clusters, related to Fig. 6 F. Mean ± SEM, three independent cultures, nonAcD: n = 34 cells, AcD: n = 32 cells. Grey dots indicate value of individual cell. Orange and cyan triangle indicate mean of each independent culture. (F) Number of homer-1 (left) and VGLUT (right) puncta per 5 μm at the AIS of nonAcD and AcD neurons. Mean ± SEM, three independent cultures for Homer-1, four independent experiments for VGLUT, Homer-1: n (nonAcD) = 74 cells, n (AcD) = 78 cells, VGLUT: n (nonAcD) = 67 cells, n (AcD) = 50 cells. Grey dots indicate value of individual cell. Orange and cyan triangle indicate mean of each independent culture. (G) Representative images of AIS-specific ECMs labeled by brevican staining at the AIS of DIV12 nonAcD. Red arrowheads indicate brevican at the AIS. Scale bar is 10 µm. Mann–Whitney test (two-sided): not significant (n.s.) P > 0.05.

Specific ECM and excitatory synapses at the AIS of AcD and nonAcD neurons. (A and B) Representative images of AIS-specific ECMs labeled by brevican staining at the AIS of DIV21 nonAcD and AcD neurons. Black arrowheads indicate brevican at the AIS. Scale bar is 10 µm. (C and D) Representative images of nonAcD and AcD neurons stained with markers for pre- and postsynaptic compartments of an excitatory synapse (VGLUT and homer-1, respectively), for AIS (TRIM46 or AnkG), and for somato-dendritic compartment (MAP2). (C) VGLUT (D) homer-1. Scale bar is 10 µm. Inset: corresponding zoom-ins of white dashed rectangular. Scale bar is 5 µm. Black arrowheads indicate corresponding markers for pre- and postexcitatory synapse. (E) Distance between co-localized gephyrin and VGAT clusters, related to Fig. 6 F. Mean ± SEM, three independent cultures, nonAcD: n = 34 cells, AcD: n = 32 cells. Grey dots indicate value of individual cell. Orange and cyan triangle indicate mean of each independent culture. (F) Number of homer-1 (left) and VGLUT (right) puncta per 5 μm at the AIS of nonAcD and AcD neurons. Mean ± SEM, three independent cultures for Homer-1, four independent experiments for VGLUT, Homer-1: n (nonAcD) = 74 cells, n (AcD) = 78 cells, VGLUT: n (nonAcD) = 67 cells, n (AcD) = 50 cells. Grey dots indicate value of individual cell. Orange and cyan triangle indicate mean of each independent culture. (G) Representative images of AIS-specific ECMs labeled by brevican staining at the AIS of DIV12 nonAcD. Red arrowheads indicate brevican at the AIS. Scale bar is 10 µm. Mann–Whitney test (two-sided): not significant (n.s.) P > 0.05.

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