PMC

AKymographs and schematic representations of time‐lapse recordings of the proximal axon, for different time points (day 7; day 13) and conditions (control; Centrinone‐B). Scale bar = 5 µm.BQuantifications of the percentage of neurons exhibiting uniform, or non‐uniform comet orientations in the anterograde direction in the proximal axon at day 7. n = 13–14 cells in four independent experiments.CQuantifications of the percentage of neurons exhibiting uniform, or non‐uniform comet orientations in the anterograde direction in the proximal axon at day 13. n = 12–17 cells in four independent experiments.DQuantifications of the number of comets per minute moving in the anterograde direction in the proximal axon at day 7 and day 13. n = 12–17 cells in four independent experiments.EQuantifications of the number of comets per minute moving in the retrograde direction in the proximal axon at day 7 and day 13. n = 12–17 cells in four independent experiments.FKymographs and schematic representations of time‐lapse recordings of the dendrite, for different time points (day 7; day 13) and conditions (control; Centrinone‐B). Scale bar = 5 µm.GQuantifications of the percentage of neurons exhibiting uniform, or non‐uniform comet orientations in the anterograde direction in the dendrite at day 7. n = 12–13 cells in four independent experiments.HQuantifications of the percentage of neurons exhibiting uniform, or non‐uniform comet orientations in the anterograde direction in the dendrite at day 13. n = 13–16 cells in four independent experiments.IQuantifications of the number of comets per minute moving in the anterograde direction in the dendrite at day 7 and day 13. n = 12–16 cells in four independent experiments.JQuantifications of the number of comets per minute moving in the retrograde direction in the dendrite at day 7 and day 13. n = 12–16 cells in four independent experiments.KKymographs and schematic representations of time‐lapse recordings of the proximal axon following LS, for different time points (day 7; day 13) and conditions (control; Centrinone‐B). Red line and red arrowhead denote location and time of LS. Scale bar = 5 µm.LQuantifications of the percentage of neurons exhibiting uniform, or non‐uniform comet orientations in the anterograde direction in the proximal axon following LS at day 7. n = 20–21 neurons in three independent experiments.MQuantifications of the percentage of neurons exhibiting uniform, or non‐uniform comet orientations in the anterograde direction in the proximal axon following LS at day 13. n = 22–25 neurons in three independent experiments.NQuantifications of the number of comets per minute moving in the anterograde direction in the proximal axon following LS at day 7 and day 13. n = 20–25 cells in three independent experiments.OQuantifications of the number of comets per minute moving in the retrograde direction in the proximal axon following LS at day 7 and day 13. n = 20–25 cells in three independent experiments.PKymographs and schematic representations of time‐lapse recordings of the dendrite following LS, for different time points (day 7; day 13) and conditions (control; Centrinone‐B). Red line and red arrowhead denote location and time of LS. Scale bar = 5 µm.QQuantifications of the number of comets per minute moving in the anterograde direction in the dendrite following LS at day 7 and day 13. n = 20–26 cells in three independent experiments.RQuantifications of the number of comets per minute moving in the retrograde direction in the dendrite following LS at day 7 and day 13. n = 20–26 cells in three independent experiments.SQuantifications of the percentage of neurons exhibiting uniform, or non‐uniform comet orientations in the anterograde direction in the distal axon at day 13 for different conditions (control; Centrinone‐B addition post‐differentiation at day 5). n = 21–23 cells in three independent experiments.TQuantifications of the number of comets per minute pointing in the anterograde direction in the distal axon at day 13 for different conditions (control; Centrinone‐B addition post‐differentiation at day 5). n = 21–23 cells in two independent experiments.UQuantifications of the number of comets per minute pointing in the retrograde direction in the distal axon at day 13 for different conditions (control; Centrinone‐B addition post‐differentiation at day 5). n = 21–23 cells in two independent experiments.
Data information: Data represent mean ± SEM. Chi‐square test (B, C, G, H, L, M, S), unpaired t‐test (D, E, I, J, N, O, Q, R, T, U); **P < 0.005, *P < 0.05, ns P ≥ 0.05.

Cross-linked number of interconnected pores along the specimen height direction. (a) Cross-linked number spatial distribution of PAC-5; (b) Cross-linked number of PAC-5 along the specimen height direction. (c) Cross-linked number of the spatial distribution of PAC-10; (d) Cross-linked number of PAC-10 along the specimen height direction; (e) Cross-linked number of the spatial distribution of PAC-13(1); (f) Cross-linked number of PAC-13(1) along the specimen height direction; (g) Cross-linked number of the spatial distribution of PAC-13(2); (h) Cross-linked number of PAC-13(2) along the specimen height direction (i) Cross-linked number spatial distribution of PAC-13(3); (j) Cross-linked number of PAC-13(3) along the specimen height direction.

AExample stills from a spinning‐disk time‐lapse recording of a neurite transfected with mRFP and GFP‐MT+TIP. The top panel is a still of a neurite in mRFP, showing neurite morphology. The other panels show moving GFP‐MT+TIP comets pointing in either an anterograde direction (green arrowheads) or retrograde direction (blue arrowheads). P indicates the proximal direction and D the distal direction of the neurite. Timestamp in minutes:seconds given on bottom right. Scale bar = 5 µm.BKymographs and schematic representations of time‐lapse recordings of the distal axon as shown in (A), for different time points (day 7; day 13) and conditions (control; Centrinone‐B). Scale bar = 5 µm.CQuantifications of the percentage of neurons exhibiting uniform, or non‐uniform comet orientations in the anterograde direction in the distal axon at day 7. n = 15–17 cells in four independent experiments.DQuantifications of the number of comets per minute moving in the anterograde direction in the distal axon at day 7. n = 15–17 cells in four independent experiments.EQuantifications of the number of comets per minute moving in the retrograde direction in the distal axon at day 7. n = 15–17 cells in four independent experiments.FQuantifications of the percentage of neurons exhibiting uniform, or non‐uniform comet orientations in the anterograde direction in the distal axon at day 13. n = 11–18 cells in four independent experiments.GQuantifications of the number of comets per minute moving in the anterograde direction in the distal axon at day 13. n = 11–18 cells in four independent experiments.HQuantifications of the number of comets per minute moving in the retrograde direction in the distal axon at day 13. n = 11–18 cells in four independent experiments.ISchematic representation of microtubule laser‐severing (LS) experiments.JExample stills from a spinning‐disk time‐lapse recording of a neurite transfected with mRFP and GFP‐MT+TIP. Red line denotes the location of LS. Scale bar = 5 µm.KKymographs and schematic representations of time‐lapse recordings of the distal axon as shown in (J) following LS, for different time points (day 7; day 13) and conditions (control; Centrinone‐B). Red line and red arrowhead denote location and time of LS. Scale bar = 5 µm.LQuantifications of the percentage of neurons exhibiting uniform, or non‐uniform comet orientations in the anterograde direction in the distal axon following LS at day 7. n = 20–22 neurons in three independent experiments.MQuantifications of the number of comets per minute moving in the anterograde direction in the distal axon following LS at day 7. n = 20–22 cells in three independent experiments.NQuantifications of the number of comets per minute moving in the retrograde direction in the distal axon following LS at day 7. n = 20–22 cells in three independent experiments.OQuantifications of the percentage of neurons exhibiting uniform, or non‐uniform comet orientations in the anterograde direction in the distal axon following LS at day 13. n = 24–27 neurons in three independent experiments.PQuantifications of the number of comets per minute moving in the anterograde direction in the distal axon following LS at day 13. n = 24–27 cells in three independent experiments.QQuantifications of the number of comets per minute moving in the retrograde direction in the distal axon following LS at day 13. n = 24–27 cells in three independent experiments.RSchematic representation of the proposed orientation of microtubules in the distal axon in control conditions, and following Centrinone‐B treatment.
Data information: Data represent mean ± SEM. Chi‐square test (C, F, L, O), unpaired t‐test (D, E, G, H, M, N, P, Q); **P < 0.005, *P < 0.05, ns P ≥ 0.05.

a, Number of DEGs plotted against the total number of cells within the control and treatment groups. Each dot represents a DGE comparison of a cell type. b-d, Cumulative distributions of the calculated effect size (b), −log10(adj. p-value) (c) and log2 fold change values (d). Distributions are shown separately for ACC, REJ and AGE DGE. Vertical lines indicate the cutoffs applied throughout the study. e, Summary of ACC DGE results. Each cell type that has at least 50 cells in IY and HY is studied in the context of ACC and hence shown. From top to bottom: control and treatment sample sizes indicated separately for AGE and ACC, the number of genes differentially expressed in AGE and ACC, overlaps between AGE and ACC. Overlaps are normalized by the number of DEGs in the union of ACC and AGE DEGs. f, Summary of REJ DGE results. Each cell type that has at least 50 cells in IA and HA is studied in the context of REJ and hence shown. From top to bottom: control and treatment sample sizes indicated separately for AGE and REJ, the number of genes differentially expressed in AGE and REJ, overlaps between AGE and REJ. Overlaps are normalized by the number of DEGs in the union of REJ and AGE DEGs. g, Percent of DEGs that change in the same direction with AGE and ACC are plotted against the total number of DEGs within AGE and ACC for each comparison. Percentages are based on the union of DEGs as defined in (e) h, Percent of DEGs that change in the opposite direction with AGE and REJ are plotted against the total number of DEGs within AGE and REJ for each comparison. Percentages are based on the union of DEGs as defined in (f). i, Fraction of DEGs changing in the same direction with AGE and ACC plotted against the fraction of DEGs changing in the opposite direction with AGE and ACC. Each dot represents a cell type of the study. Colored area indicates where more DEGs change in the same direction than in the opposite direction. j, Fraction of DEGs changing in the same direction with AGE and REJ plotted against the fraction of DEGs changing in the opposite direction with AGE and REJ. Each dot represents a cell type of the study. Colored area indicates where more DEGs change in the opposite direction than in the same direction.