Figure S1.
AcD neuron identification and examples of atypical sequence of events during AcD neuron development. (A) Schematic of AcD neuron where the point of axon start and end of soma are not parallel. Black dashed line indicates the edge of soma; orange dashed line indicates the border of axon start; red dashed line indicates the axis perpendicularly extended from the center of axon start. Related to Fig. 1 B. (B) Histogram of axon distance of AcD neurons. The median of axon distance is 7.75 µm (indicated by box plot). n = 800 cells. (C) Scatter plot of AcD neuron axon distance versus stem dendrite (Stem D) diameter. n = 800 cells. (D) Representative confocal images of nonAcD neurons and AcD neurons with different axon distance. White arrowhead indicates nonAcD neurons, orange arrowhead indicates AcD neurons. Scale bar is 20 µm. (E) AIS diameter measurement. Mean ± SEM, 4 independent cultures, nonAcD: n = 46 cells, AcD: n = 62 cells. Grey dot indicates value of individual cell. Cyan and orange triangles indicate mean of each experiment. Mann–Whitney test (two-sided): not significant (n.s.) P > 0.05. (F) Quantification of AcD neuron percentage in dissociated culture at different age. Grey dot indicates the percentage of individual images, orange triangle indicates mean of each experiment. Mean ± SEM, three independent cultures for each age, DIV5: n = 6 coverslips, DIV7: n = 6 coverslips, DIV21: n = 6 coverslips. One-way ANOVA with Tukey’s multiple comparisons test, no significance (n.s.) P > 0.05, *P < 0.05. (G) Top row: Representative images from a time-lapse recording of AcD neuron developing a precursor neurite as axon and collateral as AcD (majority of cases). Related to Fig. 1 C and Video 2. Bottom row: Representative images from time-lapse recording of AcD neuron growing a precursor neurite as AcD and collateral as axon (atypical sequence). Related to Video 3 A. Orange arrowhead indicates the precursor neurite of AcD neuron. White arrowhead indicates axon. Yellow and pink arrowhead indicates AcD and stem dendrite, respectively. Dendrites and the AIS is labeled by poststaining with MAP2 and AnkG, respectively. Scale bar is 20 µm. (H) Representative images from a time-lapse recording of an AcD neuron developing an axon and AcD via bifurcation of the precursor neurite’s growth cone. Related to Fig. 1 D and Video 3 B. Orange arrowhead indicates precursor neurite. White arrowhead indicates axon. Yellow and pink arrowhead indicates AcD and stem dendrite respectively. Cell body with dendrites and the AIS are labeled with MAP2 and AnkG antibody, respectively. Scale bar is 20 µm. (I) Schematic of neurofascin antibody live-labeling assay.

AcD neuron identification and examples of atypical sequence of events during AcD neuron development. (A) Schematic of AcD neuron where the point of axon start and end of soma are not parallel. Black dashed line indicates the edge of soma; orange dashed line indicates the border of axon start; red dashed line indicates the axis perpendicularly extended from the center of axon start. Related to Fig. 1 B. (B) Histogram of axon distance of AcD neurons. The median of axon distance is 7.75 µm (indicated by box plot). n = 800 cells. (C) Scatter plot of AcD neuron axon distance versus stem dendrite (Stem D) diameter. n = 800 cells. (D) Representative confocal images of nonAcD neurons and AcD neurons with different axon distance. White arrowhead indicates nonAcD neurons, orange arrowhead indicates AcD neurons. Scale bar is 20 µm. (E) AIS diameter measurement. Mean ± SEM, 4 independent cultures, nonAcD: n = 46 cells, AcD: n = 62 cells. Grey dot indicates value of individual cell. Cyan and orange triangles indicate mean of each experiment. Mann–Whitney test (two-sided): not significant (n.s.) P > 0.05. (F) Quantification of AcD neuron percentage in dissociated culture at different age. Grey dot indicates the percentage of individual images, orange triangle indicates mean of each experiment. Mean ± SEM, three independent cultures for each age, DIV5: n = 6 coverslips, DIV7: n = 6 coverslips, DIV21: n = 6 coverslips. One-way ANOVA with Tukey’s multiple comparisons test, no significance (n.s.) P > 0.05, *P < 0.05. (G) Top row: Representative images from a time-lapse recording of AcD neuron developing a precursor neurite as axon and collateral as AcD (majority of cases). Related to Fig. 1 C and Video 2. Bottom row: Representative images from time-lapse recording of AcD neuron growing a precursor neurite as AcD and collateral as axon (atypical sequence). Related to Video 3 A. Orange arrowhead indicates the precursor neurite of AcD neuron. White arrowhead indicates axon. Yellow and pink arrowhead indicates AcD and stem dendrite, respectively. Dendrites and the AIS is labeled by poststaining with MAP2 and AnkG, respectively. Scale bar is 20 µm. (H) Representative images from a time-lapse recording of an AcD neuron developing an axon and AcD via bifurcation of the precursor neurite’s growth cone. Related to Fig. 1 D and Video 3 B. Orange arrowhead indicates precursor neurite. White arrowhead indicates axon. Yellow and pink arrowhead indicates AcD and stem dendrite respectively. Cell body with dendrites and the AIS are labeled with MAP2 and AnkG antibody, respectively. Scale bar is 20 µm. (I) Schematic of neurofascin antibody live-labeling assay.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal