Figure 5.
AcD neurons do not differ in pan-Nav1, AnkG, and neurofascin abundancies at the AIS from nonAcD neurons but contain fewer cisternal organelles. (A) Top row: Representative images of DIV21 nonAcD (left) and AcD (right) neuron-labelled with MAP2, AIS-specific MAP TRIM46, and pan-Nav1 for sodium channels. White dashed rectangle indicates the AIS. Scale bar is 10 µm. Bottom row: corresponding zoom-ins to white dashed rectangle in upper row. Scale bar is 5 µm. (B) Top row: Representative images of DIV21 nonAcD (left) and AcD (right) neuron labelled with MAP2 and AIS specific scaffolding protein AnkG. White dashed rectangle indicates the AIS. Scale bar is 10 µm. bottom row: corresponding zoom-ins to white dashed rectangle in Upper row. Scale bar is 5 µm. (C) Top row: Representative images of DIV21 nonAcD (left) and AcD (right) neuron labelled with MAP2 and AIS specific membrane protein neurofascin. White dashed rectangle indicates the AIS. Scale bar is 10 µm. Bottom row: corresponding zoom-ins to white dashed rectangle in Upper row. Scale bar is 5 µm. (D) Top row: Representative images of DIV21 nonAcD (left) and AcD (right) neuron labeled with MAP2, AIS-specific scaffolding protein AnkG, and synaptopodin (synpo) for cisternal organelles. White dashed rectangle indicates the AIS. Scale bar is 10 µm. Bottom row: corresponding zoom-ins to white dashed rectangle in upper row. Scale bar is 5 µm. (E–G) Quantification of fluorescent intensity at the AIS of nonAcD and AcD neurons for sodium channels (E), AnkG (F), and neurofascin (G). Mean ± SEM, three independent cultures for each AIS protein, pan-Nav1: n (nonAcD) = 15 cells, n (AcD) = 19 cells, AnkG: n (nonAcD) = 41 cells, n (AcD) = 38 cells, neurofascin: n (nonAcD) = 32 cells, n (AcD) = 36 cells. Grey dot indicates value of individual cell. Orange and cyan triangle indicate mean of each independent culture. (H) Quantification of synpo cluster density (left) and size (right) at the AIS of nonAcD and AcD neurons. Mean ± SEM, three independent cultures, synpo density: n (nonAcD) = 91 cells, n (AcD) = 67 cells, cluster size: n (nonAcD) = 48 cells, n (AcD) = 54 cells. Grey dot indicates the value of individual cell. Orange and cyan triangles indicate mean of each independent culture. Mann–Whitney test (two-sided): not significant (n.s.) P > 0.05, *P < 0.05, **P < 0.01. More detailed statistical information see Data S1.

AcD neurons do not differ in pan-Nav1, AnkG, and neurofascin abundancies at the AIS from nonAcD neurons but contain fewer cisternal organelles. (A) Top row: Representative images of DIV21 nonAcD (left) and AcD (right) neuron-labelled with MAP2, AIS-specific MAP TRIM46, and pan-Nav1 for sodium channels. White dashed rectangle indicates the AIS. Scale bar is 10 µm. Bottom row: corresponding zoom-ins to white dashed rectangle in upper row. Scale bar is 5 µm. (B) Top row: Representative images of DIV21 nonAcD (left) and AcD (right) neuron labelled with MAP2 and AIS specific scaffolding protein AnkG. White dashed rectangle indicates the AIS. Scale bar is 10 µm. bottom row: corresponding zoom-ins to white dashed rectangle in Upper row. Scale bar is 5 µm. (C) Top row: Representative images of DIV21 nonAcD (left) and AcD (right) neuron labelled with MAP2 and AIS specific membrane protein neurofascin. White dashed rectangle indicates the AIS. Scale bar is 10 µm. Bottom row: corresponding zoom-ins to white dashed rectangle in Upper row. Scale bar is 5 µm. (D) Top row: Representative images of DIV21 nonAcD (left) and AcD (right) neuron labeled with MAP2, AIS-specific scaffolding protein AnkG, and synaptopodin (synpo) for cisternal organelles. White dashed rectangle indicates the AIS. Scale bar is 10 µm. Bottom row: corresponding zoom-ins to white dashed rectangle in upper row. Scale bar is 5 µm. (E–G) Quantification of fluorescent intensity at the AIS of nonAcD and AcD neurons for sodium channels (E), AnkG (F), and neurofascin (G). Mean ± SEM, three independent cultures for each AIS protein, pan-Nav1: n (nonAcD) = 15 cells, n (AcD) = 19 cells, AnkG: n (nonAcD) = 41 cells, n (AcD) = 38 cells, neurofascin: n (nonAcD) = 32 cells, n (AcD) = 36 cells. Grey dot indicates value of individual cell. Orange and cyan triangle indicate mean of each independent culture. (H) Quantification of synpo cluster density (left) and size (right) at the AIS of nonAcD and AcD neurons. Mean ± SEM, three independent cultures, synpo density: n (nonAcD) = 91 cells, n (AcD) = 67 cells, cluster size: n (nonAcD) = 48 cells, n (AcD) = 54 cells. Grey dot indicates the value of individual cell. Orange and cyan triangles indicate mean of each independent culture. Mann–Whitney test (two-sided): not significant (n.s.) P > 0.05, *P < 0.05, **P < 0.01. More detailed statistical information see Data S1.

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